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Product listing: PSMA6 Antibody, UniProt ID P60900 #2459 to UCHL1 Antibody, UniProt ID P09936 #3524

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: The 20S proteasome is the major proteolytic enzyme complex involved in intracellular protein degradation. It consists of four stacked rings, each with seven distinct subunits. The two outer layers are identical rings composed of α subunits (called PSMAs), and the two inner layers are identical rings composed of β subunits. While the catalytic sites are located on the β rings (1-3), the α subunits are important for assembly and as binding sites for regulatory proteins (4). Seven different α and ten different β proteasome genes have been identified in mammals (5). PA700, PA28, and PA200 are three major protein complexes that function as activators of the 20S proteasome. PA700 binds polyubiquitin with high affinity and associates with the 20S proteasome to form the 26S proteasome, which preferentially degrades poly-ubiquitinated proteins (1-3). The proteasome has a broad substrate spectrum that includes cell cycle regulators, signaling molecules, tumor suppressors, and transcription factors. By controlling the degradation of these intracellular proteins, the proteasome functions in cell cycle regulation, cancer development, immune responses, protein folding, and disease progression (6-9).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: The 26S proteasome is a highly abundant proteolytic complex involved in the degradation of ubiquitinated substrate proteins. It consists largely of two sub-complexes, the 20S catalytic core particle (CP) and the 19S/PA700 regulatory particle (RP) that can cap either end of the CP. The CP consists of two stacked heteroheptameric β-rings (β1-7) that contain three catalytic β-subunits and are flanked on either side by two heteroheptameric α-rings (α1-7). The RP includes a base and a lid, each having multiple subunits. The base, in part, is composed of a heterohexameric ring of ATPase subunits belonging to the AAA (ATPases Associated with diverse cellular Activities) family. The ATPase subunits function to unfold the substrate and open the gate formed by the α-subunits, thus exposing the unfolded substrate to the catalytic β-subunits. The lid consists of ubiquitin receptors and DUBs that function in recruitment of ubiquitinated substrates and modification of ubiquitin chain topology (1,2). Other modulators of proteasome activity, such as PA28/11S REG, can also bind to the end of the 20S CP and activate it (1,2).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: The 26S proteasome is a highly abundant proteolytic complex involved in the degradation of ubiquitinated substrate proteins. It consists largely of two sub-complexes, the 20S catalytic core particle (CP) and the 19S/PA700 regulatory particle (RP) that can cap either end of the CP. The CP consists of two stacked heteroheptameric β-rings (β1-7) that contain three catalytic β-subunits and are flanked on either side by two heteroheptameric α-rings (α1-7). The RP includes a base and a lid, each having multiple subunits. The base, in part, is composed of a heterohexameric ring of ATPase subunits belonging to the AAA (ATPases Associated with diverse cellular Activities) family. The ATPase subunits function to unfold the substrate and open the gate formed by the α-subunits, thus exposing the unfolded substrate to the catalytic β-subunits. The lid consists of ubiquitin receptors and DUBs that function in recruitment of ubiquitinated substrates and modification of ubiquitin chain topology (1,2). Other modulators of proteasome activity, such as PA28/11S REG, can also bind to the end of the 20S CP and activate it (1,2).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: PSMD10/Gankyrin is an ankyrin-repeat chaperone protein that is involved in the assembly of the 19S regulatory particle of the proteasome (1). Reasearch studies have demonstrated that PSMD10 is oncogenic and is overexpressed in hepatocullar carcinomas (2). Investigators believe the oncogenicity of PSMD10 may be linked to its ability to bind and regulate the stability and activity of pRB, CDK4, MDM2, and RelA (2-5).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunoprecipitation, Western Blotting

Background: The 26S proteasome is a highly abundant proteolytic complex involved in the degradation of ubiquitinated substrate proteins. It consists largely of two sub-complexes, the 20S catalytic core particle (CP) and the 19S/PA700 regulatory particle (RP) that can cap either end of the CP. The CP consists of two stacked heteroheptameric β-rings (β1-7) that contain three catalytic β-subunits and are flanked on either side by two heteroheptameric α-rings (α1-7). The RP includes a base and a lid, each having multiple subunits. The base, in part, is composed of a heterohexameric ring of ATPase subunits belonging to the AAA (ATPases Associated with diverse cellular Activities) family. The ATPase subunits function to unfold the substrate and open the gate formed by the α-subunits, thus exposing the unfolded substrate to the catalytic β-subunits. The lid consists of ubiquitin receptors and DUBs that function in recruitment of ubiquitinated substrates and modification of ubiquitin chain topology (1,2). Other modulators of proteasome activity, such as PA28/11S REG, can also bind to the end of the 20S CP and activate it (1,2).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: RING-box protein 1 (RBX1 or ROC1) is an essential component of two distinct but structurally related E3 ubiquitin ligase complexes, the SCF complex and the CBC (VHL) complex (1). RBX1 mediates the neddylation of CUL1, which activates SCF E3 ligase by facilitating the ubiquitin transfer from E2 to substrates (2-4). The RING finger domain of RBX1 is required for ubiquitin ligation (5). Two evolutionarily conserved mammalian RBX family members, RBX1/ROC1 and RBX2/ROC2/SAG, have been identified (5). RBX1 is constitutively expressed and binds to CUL2/VHL, while stress-inducible RBX2 binds to CUL5/SOCS (6).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse

Application Methods: Immunoprecipitation, Western Blotting

Background: RING finger and CHY1 zinc finger domain-containing protein 1 (RCHY1) is a newly identified RING-H2-type protein-ubiquitin E3 ligase that is expressed as multiple isoforms generated through alternative splicing of mRNA transcripts (1,2). Notably, RCHY1 is a p53-regulated gene and multiple studies have shown that RCHY1 directly binds p53 to promote its ubiquitin-dependent proteasomal degradation, ultimately leading to repression of p53 growth suppressive transcriptional activity (1,3). RCHY1 is also likely to promote deregulated cell proliferation by targeting the CDK inhibitor, p27 Kip1, for proteasomal degradation (4). In agreement with its growth promoting activity, RCHY1 overexpression has been linked to cancer of the lung (5,6) and prostate (7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: S5a (PSMD4) is a subunit of the 19S regulatory proteasome complex functioning in ubiquitinated-protein targeting and degradation (1). S5a contains two polyubiquitin binding motifs (UIM) that bind multiubiquitin chains by hydrophobic interaction (2,3). In addition to ubiquitin, the UIM of S5a shows high affinity to a ubiquitin-like domain present in many proteins. S5a binds to these types of proteins directly and mediates their targeting to the proteasome for degradation (4,5).

$111
20 µl
$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry), Western Blotting

Background: Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2 and then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (1-3). Combinatorial interactions of different E2 and E3 proteins result in substrate specificity (4). Recent data suggests that activated E2 associates transiently with E3, and the dissociation is a critical step for ubiquitination (5). S phase kinase-associated protein 1 (Skp1) is a critical scaffold protein of the Skp1/CUL1/F-box (SCF) E3 ubiquitin ligase protein complex. Various F-box proteins (e.g., β-TrCP, Skp2) mediate an interaction with Skp1, via their defining and conserved domain of 40 amino acids, and with substrates to be ubiquitinated (e.g., β-catenin, p27) (4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Immunofluorescence (Immunocytochemistry), Western Blotting

Background: Members of the F-box family of proteins are characterized by the approximate 40 amino acid F-box motif named after cyclin F (1,2). F-box proteins constitute one of the four subunits of the Skp1-Cullin-F-box (SCF) ubiquitin ligase complex. The substrate specificity of SCF complexes is determined by the interchangeable F-box proteins, which act as adaptors by associating with phosphorylated substrate proteins and recruiting them to the SCF core. F-box proteins contain two fundamental domains: the F-box motif mediates binding to Skp1 and a leucine rich repeat (LRR) domain mediates substrate interactions.

$111
20 µl
$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunofluorescence (Immunocytochemistry), Western Blotting

Background: Members of the F-box family of proteins are characterized by the approximate 40 amino acid F-box motif named after cyclin F (1,2). F-box proteins constitute one of the four subunits of the Skp1-Cullin-F-box (SCF) ubiquitin ligase complex. The substrate specificity of SCF complexes is determined by the interchangeable F-box proteins, which act as adaptors by associating with phosphorylated substrate proteins and recruiting them to the SCF core. F-box proteins contain two fundamental domains: the F-box motif mediates binding to Skp1 and a leucine rich repeat (LRR) domain mediates substrate interactions.

$260
100 µl
APPLICATIONS
REACTIVITY
Mouse, Rat

Application Methods: Western Blotting

Background: The Kazal type 3 serine peptidase inhibitor (SPINK3) is a serine peptidase inhibitor expressed in the adult pancreas that protects acinar cells from trypsin-induced cell damage (1). Ingestion of milk by newborn SPINK3 knock out mice triggers autophagic cell death in pancreatic acinar cells and the complete disappearance of the pancreas (2). Mutations in the human SPINK3 homolog (known as SPINK1) are associated with chronic pancreatitis, presumably through decreased protection against trypsin-induced autophagic cell death (1,3). SPINK3 may be involved in the embryonic development of several other tissues (4).

$111
20 µl
$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunoprecipitation, Western Blotting

Background: Protein ubiquitination and deubiquitination is a reversible process catalyzed by ubiquitinating enzymes (UBEs) and deubiquitinating enzymes (1,2). Deubiquitinating enzymes (DUBs) function as ubiquitin-specific proteases and can be divided into five subfamilies based on catalytic domain structure. At least 14 members of the JAMM ubiquitin protease subfamily have been identified, including signal transducing adaptor molecule (STAM) binding protein (3). STAM-binding protein (STAMBP or AMSH) is an endosomal DUB that preferentially displays ubiquitin isopeptidase activity toward K63-linked chains, which is critically dependent upon its interaction with STAM (4,5). STAMBP interacts with the STAM adaptor protein and becomes integrated into the multivesicular body sorting machinery to help regulate endosomal trafficking and receptor tyrosine kinase stability by deubiquitining target proteins (4,6). Evidence indicates that endosomal STAMBP antagonizes the ubiquitin-dependent trafficking of EGFR to the lysosomal compartment (7).

$111
20 µl
$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Immunofluorescence (Immunocytochemistry), Immunohistochemistry (Paraffin), Immunoprecipitation, Western Blotting

Background: Small ubiquitin-related modifier 1, 2 and 3 (SUMO-1, -2 and -3) are members of the ubiquitin-like protein family (1). The covalent attachment of the SUMO-1, -2 or -3 (SUMOylation) to target proteins is analogous to ubiquitination. This post-translational modification is a reversible, multi-step process that is initiated by cleaving a precursor protein to a mature protein. Mature SUMO-1, -2 or -3 is then linked to the activating enzyme E1, conjugated to E2 and in conjunction with E3, SUMO-1, -2 or -3 is ligated to the target protein (2). Ubiquitin and the individual SUMO family members are all targeted to different proteins with diverse biological functions. Ubiquitin predominantly regulates degradation of its target (1). In contrast, SUMO-1 is conjugated to RanGAP, PML, p53 and IκB-α to regulate nuclear trafficking, formation of subnuclear structures, regulation of transcriptional activity and protein stability (3-7). SUMO-2/-3 forms poly-(SUMO) chains, is conjugated to topoisomerase II and APP, regulates chromosomal segregation and cellular responses to environmental stress, and plays a role in the progression of Alzheimer disease (8-11).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: Tripeptidyl-peptidase 2 (TPP2) is a well-conserved subtilisin-like amino peptidase that is expressed predominantly in the cytoplasmic compartment (1,2). The amino-terminal region of TPP2 harbors a catalytic triad that is characteristic of serine proteases and allows for TPP2 cleavage of tripeptides from the free amino terminus of oligopeptide substrates (3). TPP2 is a large (>5MDa) homooligomeric protease in which proteolytic activity is regulated by subunit oligomerization (4,5). While TPP2 plays a general role in amino acid homeostasis, research studies demonstrate that TPP2 is involved in MHC class I antigen presentation (6,7) and DNA-damage repair (8). TPP2 activity is required for the survival of Burkitt's lymphoma cells, suggesting a possible role for TPP2 in oncogenesis (9). Additional research studies show that TPP2 proteolytic activity is important for regulating lysosome abundance and glycolytic metabolism and that TPP2 deficiency leads to defects in adaptive immunity, innate immunity, and nervous system development (10).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: The E3 ubiquitin-protein ligase ARIH2 (TRIAD1) is an Ariadne subfamily ligase involved in the polyubiquitination of proteins designated for proteasomal degradation. The TRIAD1 nuclear protein contains an amino-terminal acidic region, a pair of RING fingers, two carboxyl-terminal coiled coil domains and a novel C6HC DRIL/IBR domain located between the RING fingers. Together, the paired RING fingers and DRIL/IBR domain form a highly conserved TRIAD (two RING fingers and DRIL) domain (1). Research studies suggest that TRIAD1 mediates both Lys48 and Lys63 protein polyubiquitination and acts as a negative regulator of myelopoiesis. TRIAD1 ubiquitin ligase inhibits myeloid cell proliferation by mediating protein ubiquitination through the ubiquitin-conjugating enzymes UbcH7 and UbcH13 (2,3). Experimental deletion of TRIAD1 in mice has a lethal effect, leading to death at the embryonic stage or later due to a severe, multi-organ inflammatory response. Results indicate that TRIAD1 binds IκBβ in dendritic cells and promotes the degradation of the NF-κB inhibitor (4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: The process of SUMO conjugation to target proteins is similar to the molecular chain of events observed with ubiquitin (1). SUMO is conjugated to target proteins through the coordinated action of the cellular SUMO conjugation machinery consisting of E1, E2, and E3 enzymes (2). The canonical SUMO E1 activating enzyme is a heterodimer consisting of SAE1 (AOS1) and UBA2 (SAE2) subunits. Mature SUMO is activated by E1 in an ATP-dependent reaction that generates adenylated SUMO, which functions as a high-energy intermediate in the formation of a thioester linkage between SUMO and Cys173 of UBA2 (3,4). SUMO is subsequently transferred from UBA2 to the SUMO E2 conjugating enzyme, UBC9 (5). Recent evidence suggests that redox regulation of UBA2 serves as a physiologic mechanism to modulate the cellular level of sumoylated target proteins (6).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: Similar to ubiquitin, NEDD8 is covalently linked to target proteins through an enzymatic cascade composed of NEDD8-specific E1 (activating)- and E2 (conjugating)-enzymes (1,2). The E2 ligase specific for NEDD8 is Ubc12 (3-5). Ubc12 forms a heterodimeric conjugate with NEDD8 in order to catalyze the transfer of NEDD8 from E1 to lysine side chains of target proteins (1,2). Well known targets of NEDD8 are cullin-based RING E3 ligases. Neddylation of cullin isoforms activates the related ubiquitin E3 complex by promoting its interaction with a cognate ubiquitin-E2 ligase (6-7). Neddylation of Cul-1 complexes containing βTrCP and SKP2 has been shown to be required for controlling the stability of important signaling targets such as IκB, NF-κB, and p27 Kip (8-10), thereby regulating cell cycle progression, signaling cascades, and developmental programming processes (11).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: Protein ubiquitination is an important posttranslational modification that regulates protein function and fate (1). Ubiquitin (Ub) can be conjugated to target proteins in either monomeric or polymeric forms. There are several different lysine residues within Ub that can be used as conjugation sites for poly-Ub chain formation. Different poly-Ub linkages mediate different functions of the target protein ranging from alterations in protein function to degradation (2). UBE2N/Ubc13 is a ubiquitin-E2-conjugating enzyme that catalyzes K63-linked poly-Ub chain formation (1,2). UBE2N forms a heterodimer with MMS2 or Uev1A to exert its E2 ligase function. The UBE2N/MMS2 and UBE2N/Uev1A heterodimers catalyze different modes of target protein ubiquitination to mediate various signaling pathways (3-5) including: DNA damage and recombination, p53 and check point control, the cell cycle (6-10), immunoreceptor signaling (11,12), and endocytosis (13). Most recently, UBE2N was shown to play an important role in inflammatory signaling by promoting K63-linked ubiquitination and activation of IKK downstream of the IL-1β receptor (14). Furthermore, interaction of UBE2N with the Triad1 E3 protein-ubiquitin ligase was shown to play an important role in myelopoiesis (15).

$111
20 µl
$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Flow Cytometry, Western Blotting

Background: Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, and then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (1-3). Combinatorial interactions of different E2 and E3 proteins result in substrate specificity (4). Recent data suggest that activated E2 associates transiently with E3, and that the dissociation is a critical step for ubiqitination (5). UBC3, the mammalian orthologue of yeast Cdc34, and UBC3B, a UBC3 family member, are E2 ubiquitin-carrier proteins. These proteins contain a conserved core domain containing a cysteine residue, which forms the thioester bond with ubiquitin (6). UBC3 in concert with the SCFSkp2 (Skp1, Cullin and F-box protein/Skp2) complex mediates cell cycle progression from G1 to S phase by targeting the CDK inhibitor p27 for proteolysis (7). UBC3B in concert with the SCFb-Trcp (Skp1, Cullin and F-box protein/b-Trcp) complex mediates degradation of b-catenin (6).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: The process of SUMO-1 conjugation is similar to that seen with ubiquitin and other forms of post-translational protein modification (1). Like ubiquitin, SUMO-1 is conjugated to its target protein by the coordinated action of ubiquitin conjugation enzymes E1, E2 and E3 (2). Ubc9 (or ube2M) is a highly conserved, 158 amino acid protein that acts as a SUMO-1 conjugating enzyme (3). Ubc9 binds to target proteins through their SUMO-1-CS (consensus sequence) domains and interacts with SUMO via the structurally conserved amino-terminal domain (3,4). Localization of Ubc9 to the nucleus and the nuclear envelope allows this enzyme to catalyze target protein sumoylation and regulate target protein nucleocytoplasmic transport and transcriptional activity (5,6). Ubc9 target proteins include a host of proteins (RAD51, RAD52, p53 and c-Jun) that regulate the cell cycle, DNA repair, and p53-dependent processes (7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry), Immunohistochemistry (Paraffin), Immunoprecipitation, Western Blotting

Background: Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the ubiquitin-activating enzyme (UBE1 or E1). The activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, and then from E2 to ubiquitin ligase E3 for final delivery to the ε-amino group of the target protein lysine residue (1-3). Combinatorial interactions of different E2 and E3 proteins result in substrate specificity (4). UBE1 has two isofoms: UBE1a is a nuclear protein of 117 kDa while UBE1b is a nuclear and cytoplasmic protein of 110 kDa (5).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Immunofluorescence (Immunocytochemistry), Western Blotting

Background: Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the ubiquitin-activating enzyme (UBE1 or E1). The activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, and then from E2 to ubiquitin ligase E3 for final delivery to the ε-amino group of the target protein lysine residue (1-3). Combinatorial interactions of different E2 and E3 proteins result in substrate specificity (4). UBE1 has two isofoms: UBE1a is a nuclear protein of 117 kDa while UBE1b is a nuclear and cytoplasmic protein of 110 kDa (5).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thioester complex with the ubiquitin-activating enzyme (E1). The activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, and then from E2 to ubiquitin ligase E3 for final delivery to the ε-amino group of the target protein lysine residue (1-3).Ubiquitin-activating enzyme E1-like protein 2/Ubiquitin-like modifier-activating enzyme 6 (UBE1L2/UBA6) is ubiquitously expressed in human tissues and functions as an E1 enzyme related to UBE1/UBA1 (40% identity at the protein level). UBE1L2/UBA6 activates both ubiquitin and the ubiquitin-like protein FAT10 through a similar ATP dependent mechanism (4-6). Like other E1 protein family members, UBE1L2/UBA6 contains a conserved ATP-binding adenylation domain and an active site cysteine residue that are critical for enzymatic function (4,5). Research studies have demonstrated that UBE1L2/UBA6 expression is essential during the early stages of embryogenesis in mice (4). Furthermore, loss of neuronal UBE1L2/UBA6 expression promotes significant defects in neuronal structure and function, which contributes to a reduction in body weight and decreased postnatal viability (7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: Protein ubiquitination requires the concerted action of the E1, E2, and E3 ubiquitin-conjugating enzymes. Ubiquitin is first activated through ATP-dependent formation of a thiol ester with ubiquitin-activating enzyme E1. The activated ubiquitin is then transferred to a thiol group of ubiquitin-carrier enzyme E2. The final step is the transfer of ubiquitin from E2 to an ε-amino group of the target protein lysine residue, which is mediated by ubiquitin-ligase enzyme E3 (1).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: Protein ubiquitination requires the concerted action of the E1, E2, and E3 ubiquitin-conjugating enzymes. Ubiquitin is first activated through ATP-dependent formation of a thiol ester with ubiquitin-activating enzyme E1. The activated ubiquitin is then transferred to a thiol group of ubiquitin-carrier enzyme E2. The final step is the transfer of ubiquitin from E2 to an ε-amino group of the target protein lysine residue, which is mediated by ubiquitin-ligase enzyme E3 (1).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Western Blotting

Background: Protein ubiquitination requires the concerted action of the E1, E2, and E3 ubiquitin-conjugating enzymes. Ubiquitin is first activated through ATP-dependent formation of a thiol ester with ubiquitin-activating enzyme E1. The activated ubiquitin is then transferred to a thiol group of ubiquitin-carrier enzyme E2. The final step is the transfer of ubiquitin from E2 to an ε-amino group of the target protein lysine residue, which is mediated by ubiquitin-ligase enzyme E3 (1).

$260
100 µl
APPLICATIONS
REACTIVITY
All Species Expected

Application Methods: Immunohistochemistry (Paraffin), Western Blotting

Background: Ubiquitin is a conserved polypeptide unit that plays an important role in the ubiquitin-proteasome pathway. Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (1-3). The ubiquitin-proteasome pathway has been implicated in a wide range of normal biological processes and in disease-related abnormalities. Several proteins such as IκB, p53, cdc25A, and Bcl-2 have been shown to be targets for the ubiquitin-proteasome process as part of regulation of cell cycle progression, differentiation, cell stress response, and apoptosis (4-7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunoprecipitation, Western Blotting

Background: The process of SUMO conjugation to target proteins is similar to the molecular chain of events observed with ubiquitin (1). SUMO is conjugated to target proteins through the coordinated action of the cellular SUMO conjugation machinery, which consists of the E1, E2, and E3 enzymes (2). The canonical SUMO E1 activating enzyme is a heterodimer consisting of Ubiquitin-like 1-activating enzyme E1A (UBLE1A, SAE1) and UBLE1B (SAE2, UBA2) subunits. Mature SUMO is activated by E1 in an ATP-dependent reaction that generates adenylated SUMO, which functions as a high-energy intermediate in the formation of a thioester linkage between SUMO and Cys173 of SAE2 (3,4). SUMO is subsequently transferred from SAE2 to the SUMO E2 conjugating enzyme UBE2I (5). Research studies indicate that UBLE1A (SAE1) is a nuclear protein and c-Myc transcriptional target whose expression is required for Myc-driven tumorigenesis (6-8).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: Protein ubiquitination and deubiquitination are reversible processes catalyzed by ubiquitinating enzymes (UBEs) and deubiquitinating enzymes (DUBs) (1,2). DUBs are categorized into 5 subfamilies: USP, UCH, OTU, MJD, and JAMM. UCHL1, UCHL3, UCHL5/UCH37, and BRCA-1-associated protein-1 (BAP1) belong to the UCH family of DUBs, which all posses a conserved catalytic domain (UCH domain) of about 230 amino acids. UCHL5 and BAP1 have unique extended C-terminal tails. UCHL1 is abundantly expressed in neuronal tissues and testes, while UCHL3 expression is more widely distributed (3,4). Although UCHL1 and UCHL3 are the most closely related UCH family members with about 53% identity, their biochemical properties differ in that UCHL1 binds monoubiquitin and UCHL3 shows dual specificity toward both ubiquitin (Ub) and NEDD8, a Ub-like molecule. In particular, UCHL3 functions as a Ub hydrolase involved in the processing of both Ub precursors and ubiquitinated substrates, generating free monomeric Ub. This is accomplished through the ability of UCHL3 to recognize and hydrolyze isopeptide bonds at the C-terminal glycine of either Ub or NEDD8 (5-7). Recent functional studies have identified UCH-L3 as a critical regulator of adipogenesis through its ability to promote IGF-IR and insulin receptor signaling (8). Furthermore, UCHL3 has been shown to promote deubiquitination, recycling, and cell surface expression of the epithelial sodium channel (9).