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Product listing: AMPK and ACC Antibody Sampler Kit #9957 to ER-Tracker™ Green (BODIPY® FL Glibenclamide) #8787

The AMPK and ACC Antibody Sampler Kit provides an economical means to investigate energy homeostasis and fatty acid synthesis within the cell. The kit contains primary and secondary antibodies to perform two Western blots with each antibody.
The Apoptosis Antibody Sampler Kit provides an economical means to evaluate the levels of inactive and active caspases. The kit contains enough primary and secondary antibodies to perform two Western blot experiments with each antibody.

Background: Apoptosis is a regulated physiological process leading to cell death. Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Initiator caspases (including 8, 9, 10 and 12) are closely coupled to proapoptotic signals. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6 and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF and lamin A, and induce apoptosis. Cytochrome c released from mitochondria is coupled to the activation of caspase-9, a key initiator caspase (1). Proapoptotic stimuli include the FasL, TNF-α, DNA damage and ER stress. Fas and TNFR activate caspases 8 and 10 (2), DNA damage leads to the activation of caspase-9 and ER stress leads to the calcium-mediated activation of caspase-12 (3). The inhibitor of apoptosis protein (IAP) family includes XIAP and survivin and functions by binding and inhibiting several caspases (4,5). Smac/Diablo, a mitochondrial protein, is released into the cytosol upon mitochondrial stress and competes with caspases for binding of IAPs. The interaction of Smac/Diablo with IAPs relieves the inhibitory effects of the IAPs on caspases (6).

The Apoptosis Antibody Sampler Kit (Mouse Specific) is designed for use with mouse samples and offers an economical means to evaluate the levels of active and inactive caspases. The kit contains enough primary and secondary antibodies to perform two Western blot experiments with each antibody.

Background: Apoptosis is a regulated physiological process leading to cell death. Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Initiator caspases (including 8, 9, 10 and 12) are closely coupled to proapoptotic signals. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6 and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF and lamin A, and induce apoptosis. Cytochrome c released from mitochondria is coupled to the activation of caspase-9, a key initiator caspase (1). Proapoptotic stimuli include the FasL, TNF-α, DNA damage and ER stress. Fas and TNFR activate caspases 8 and 10 (2), DNA damage leads to the activation of caspase-9 and ER stress leads to the calcium-mediated activation of caspase-12 (3). The inhibitor of apoptosis protein (IAP) family includes XIAP and survivin and functions by binding and inhibiting several caspases (4,5). Smac/Diablo, a mitochondrial protein, is released into the cytosol upon mitochondrial stress and competes with caspases for binding of IAPs. The interaction of Smac/Diablo with IAPs relieves the inhibitory effects of the IAPs on caspases (6).

The Apoptosis/Necroptosis Antibody Sampler Kit provides an economical means of detecting markers for apoptosis and necroptosis. The kit contains enough primary antibody to perform at least two western blot experiments.

Background: Apoptosis is a regulated physiological process leading to cell death (1,2). Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Caspases are synthesized as inactive zymogens containing a pro-domain followed by large (p20) and small subunits (p10) that are proteolytically processed in a cascade of caspase activity. Initiator caspases (including 8, 9, 10, and 12) are closely coupled to proapoptotic signals. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6, and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF, and lamin A, and induce apoptosis. Cytochrome c released from mitochondria is coupled to the activation of caspase-9, a key initiator caspase. Apoptosis induced through the extrinsic mechanisms involving death receptors in the tumor necrosis factor receptor superfamily activates caspase-8. Activated caspase-8 cleaves and activates downstream effector caspases, such as caspase-1, -3, -6, and -7. Caspase-3 is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins, such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP).Necroptosis, a regulated pathway for necrotic cell death, is triggered by a number of inflammatory signals, including cytokines in the tumor necrosis factor (TNF) family, pathogen sensors such as toll-like receptors (TLRs), and ischemic injury (3,4). Necroptosis is negatively regulated by caspase-8 mediated apoptosis in which the kinase RIP/RIPK1 is cleaved (5). Furthermore, necroptosis is inhibited by a small molecule inhibitor of RIP, necrostatin-1 (Nec-1) (6). Research studies show that necroptosis contributes to a number of pathological conditions, and Nec-1 has been shown to provide neuroprotection in models such as ischemic brain injury (7). RIP is phosphorylated at several sites within the kinase domain that are sensitive to Nec-1, including Ser14, Ser15, Ser161, and Ser166 (8). Phosphorylation drives association with RIP3, which is required for necroptosis (9-11). Mixed lineage kinase domain-like protein (MLKL) is a pseudokinase that was identified as downstream target of RIP3 in the necroptosis pathway (12). During necroptosis RIP3 is phosphorylated at Ser227, which recruits MLKL and leads to its phosphorylation at Thr357 and Ser358 (12). Knockdown of MLKL through multiple mechanisms results in inhibition of necroptosis (13). While the precise mechanism for MLKL-induced necroptosis is unclear, some studies have shown that necroptosis leads to oligomerization of MLKL and translocation to the plasma membrane, where it effects membrane integrity (14-17).

$759
30 rxns
1 Kit
The Active Ras Detection Kit provides all reagents necessary for measuring activation of Ras GTPase in the cell. GST-Raf1-RBD fusion protein is used to bind the activated form of GTP-bound Ras, which can then be immunoprecipitated with glutathione resin. Ras activation levels are then determined in western using a Ras mouse mAb.
APPLICATIONS
REACTIVITY
Human, Mouse

Application Methods: Immunoprecipitation, Western Blotting

Background: The Ras superfamily of small GTP-binding proteins (G proteins) comprise a large class of proteins (over 150 members) that can be classified into at least five families based on their sequence and functional similarities: Ras, Rho, Rab, Arf, and Ran (1-3). These small G proteins have both GDP/GTP-binding and GTPase activities and function as binary switches in diverse cellular and developmental events that include cell cycle progression, cell survival, actin cytoskeletal organization, cell polarity and movement, and vesicular and nuclear transport (1). An upstream signal stimulates the dissociation of GDP from the GDP-bound form (inactive), which leads to the binding of GTP and formation of the GTP-bound form (active). The activated G protein then goes through a conformational change in its downstream effector-binding region, leading to the binding and regulation of downstream effectors. This activation can be switched off by the intrinsic GTPase activity, which hydrolyzes GTP to GDP and releases the downstream effectors. These intrinsic guanine nucleotide exchange and GTP hydrolysis activities of Ras superfamily proteins are also regulated by guanine nucleotide exchange factors (GEFs) that promote formation of the active GTP-bound form and GTPase activating proteins (GAPs) that return the GTPase to its GDP-bound inactive form (4).

$430
100 assays
1 Kit
The Annexin V-FITC Early Apoptosis Detection Kit enables researchers to identify early apoptotic cells within a cell population. Annexin V-FITC conjugated protein binds to cell surfaces expressing phosphatidylserine, an early apotosis marker. Cells stained with propidium iodide (PI), a non-cell-permeable DNA dye, indicate necrotic cells. Cells stained with both PI and annexin V-FITC demonstrate later stage apoptosis and early necrosis. This kit provides enough reagent to perform 100 assays, based on a 250 μl assay volume.
APPLICATIONS
REACTIVITY
All Species Expected

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry)

The Malachite Green Phosphate Detection Kit is a convenient and sensitive, single-step free-phosphate determination kit that can be used for measuring phosphate released during enzymatic phosphatase assays.
The BAF Complex Antibody Sampler Kit II provides an economical means of detecting levels of various BAF complex components. The kit contains enough primary antibodies to perform at least two western blot experiments.
$29
5 x 1ml
5 ml
EDTA (Ethylenediaminetetraacetic acid) is a common laboratory chelating agent of divalent cations, such as Ca2+ and Mg2+. Ultrapure 0.5 M EDTA, pH 8.0 from Cell Signaling Technology contains no detectable DNase, RNase, or protease activity. The convenient 1 ml vials reduce the likelihood of contamination that can occur with larger volume containers. It is suitable for use in molecular biology or protein biochemistry applications that require the chelation of divalent metal cations.This product is used in our SimpleChip® chromatin immunoprecipitation (ChIP) assays to stop the metal-dependant enzymatic digestion of cross-linked DNA by micrococcal nuclease once the reaction is complete. It can be added to cell lysis buffers for use as a metalloprotease inhibitor. Working concentrations are typically 1-5 mM in this application.
Adenosine-5'-triphosphate (ATP) supplied as a 10 mM solution in doubly distilled water as a disodium salt.
Dithiothreitol (DTT) from Cell Signaling Technology is offered in a convenient 192.8 mg lyophilized format, allowing for preparation of a fresh stock solution. This DTT reagent contains no detectable DNase or RNase activity and is suitable for use in molecular biology or protein biochemistry applications that require reduction of protein disulfide bonds.SDS-PAGE sample buffers are routinely supplemented with 10-50 mM DTT to cleave protein disulfide bonds. Lower concentrations of DTT are routinely used to stabilize enzymes or other proteins that posses free sulfhydryl groups, which is useful in chromatin immunoprecipitation (ChIP) assays.
$64
10 ml
Premium quality normal goat serum for use as blocking reagent and antibody diluent for chromogenic and fluorescent immunohistochemical and immunocytochemical assays. Typically, normal serum from the same species as the secondary antibody is used in the blocking and diluent buffers.
APPLICATIONS

Application Methods: Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry), Immunofluorescence (Paraffin), Immunohistochemistry (Frozen), Immunohistochemistry (Paraffin)

$219
1 ml
This Phosphatase Inhibitor Cocktail is used to prevent dephosphorylation of phosphorylated proteins from active serine/threonine and tyrosine phosphatases present in whole cell extract. The 100X cocktail is a colorless to light yellow liquid.
APPLICATIONS

Application Methods: Western Blotting

$61
500 ml
Ponceau S Staining Solution is supplied as ready to use. This product is recommended for rapid and reversible protein staining on nitrocellulose or PVDF membranes. This staining technique is often utilized to confirm protein electrotransfer in Western blotting assays prior to antibody-based detection.
APPLICATIONS
REACTIVITY
All Species Expected

Application Methods: Western Blotting

$187
10 ml
APPLICATIONS

Application Methods: Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry), Immunofluorescence (Paraffin)

$250
1 ml
When diluted in lysis buffer to a final concentration of 1X the Protease/Phosphatase Inhibitor Cocktail prevents protein degradation and dephosphorylation by endogenous proteases and phosphatases present in the whole cell extract. The 100X cocktail is a clear light yellow to light green liquid.
APPLICATIONS
REACTIVITY
All Species Expected

Application Methods: Western Blotting

The Resazurin Cell Viability Kit is a fluorescent assay that detects cellular metabolic activity. The blue nonfluorescent resazurin reagent is reduced to highly fluorescent resorufin by dehydrogenase enzymes in metabolically active cells. This conversion only occurs in viable cells and thus, the amount of resorufin produced is proportional to the number of viable cells in the sample. The resorufin formed in the assay can be quantified by measuring the relative fluorescence units (RFU) using a fluorometer (Ex=530-570 nm, Em=590-620 nm).
$65
1 ml
$296
5 x 1ml
5 ml
The SimpleChIP® Universal qPCR Master Mix is an optimized 2X reaction mix for real-time qPCR detection and quantitation of target DNA sequences using the SYBR/FAM channel of most real-time qPCR instruments. It contains Hot Start Taq DNA Polymerase and has been formulated with a unique passive reference dye that is compatible across a variety of instrument platforms (including those that require a high or low ROX reference signal). It also features dUTP for carryover prevention and a non-fluorescent, visible dye to monitor reaction setup. This dye does not spectrally overlap with fluorescent dyes used for qPCR and will not interfere with real-time detection.This product is provided in 1 ml volumes sufficient for preparation of 100 qPCR reactions, and is compatible with both enzymatic and sonication-fragmented DNA samples from SimpleChIP® enzymatic and sonication ChIP kits. This master mix formulation is supplied at 2X concentration and contains all PCR components required for amplification and quantitation of DNA, except primers and a DNA template.
REACTIVITY
All Species Expected

Background: Dye-based quantitative PCR (qPCR) uses real-time fluorescence of a double-stranded DNA (dsDNA) binding dye, most commonly SYBR® Green I, to measure DNA amplification as it occurs during each cycle of PCR. At a point where the fluorescence signal is confidently detected over the background fluorescence, a quantification cycle, or CT value, can be determined. CT values can be used to evaluate relative target abundance between two or more samples, or to calculate absolute target quantities in reference to an appropriate standard curve, derived from a series of known dilutions. qPCR is commonly used to detect and quantify target genes in genomic DNA that is enriched by chromatin immunoprecipitation (ChIP).

$208
20 assays
1 Kit
The Cell Fractionation Kit is designed to provide a fast and efficient way of separating cultured cells into three distinct fractions: cytoplasmic, membrane/organelle, and nuclear/cytoskeletal. These fractions can then be analyzed by SDS-PAGE and western blotting. The kit includes enough buffer for 20 assays.
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

The Cell Cycle/Checkpoint Antibody Sampler Kit provides a fast and economical means of evaluating multiple proteins involved in the cell cyle and checkpoint control. The kit contains enough primary and secondary antibody to perform four Western blot experiments.

Background: The cell division cycle demands accuracy to avoid the accumulation of genetic damage. This process is controlled by molecular circuits called "checkpoints" that are common to all eukaryotic cells (1). Checkpoints monitor DNA integrity and cell growth prior to replication and division at the G1/S and G2/M transitions, respectively. The cdc2-cyclin B kinase is pivotal in regulating the G2/M transition (2,3). Cdc2 is phosphorylated at Thr14 and Tyr15 during G2-phase by the kinases Wee1 and Myt1, rendering it inactive. The tumor suppressor protein retinoblastoma (Rb) controls progression through the late G1 restriction point (R) and is a major regulator of the G1/S transition (4). During early and mid G1-phase, Rb binds to and represses the transcription factor E2F (5). The phosphorylation of Rb late in G1-phase by CDKs induces Rb to dissociate from E2F, permitting the transcription of S-phase-promoting genes. In vitro, Rb can be phosphorylated at multiple sites by cdc2, cdk2, and cdk4/6 (6-8). DNA damage triggers both the G2/M and the G1/S checkpoints. DNA damage activates the DNA-PK/ATM/ATR kinases, which phosphorylate Chk at Ser345 (9), Chk2 at Thr68 (10) and p53 (11). The Chk kinases inactivate cdc25 via phosphorylation at Ser216, blocking the activation of cdc2.

$118
10 western blots
100 µl
Caspase-3 Control Cell Extracts (Jurkat Untreated): Untreated Jurkat cells are lysed in Chaps cell extract buffer and a cytoplasmic fraction is generated to serve as a negative control for caspase cleavage. Supplied in SDS sample buffer.Caspase-3 Control Cell Extracts (Jurkat +Cytochrome c): Untreated Jurkat cells are lysed in Chaps cell extract buffer and a cytoplasmic fraction is generated. Extracts are treated with cytochrome c in vitro to generate a positive control for caspase cleavage. Supplied in SDS sample buffer.
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins, such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (1). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments. Cleavage of caspase-3 requires the aspartic acid residue at the P1 position (2).

$118
10 western blots
150 µl
Nonphosphorylated p44/42 MAPK (Erk1/2) Control Cell Extracts: Total cell extracts from Jurkat cells treated with U0126 (MEK1/2 Inhibitor) #9903 at 10 μM for 1 hour, to serve as a negative control. Supplied in SDS Sample Buffer.Phosphorylated p44/42 MAPK (Erk1/2) Control Cell Extracts: Total cell extracts from Jurkat cells treated with TPA #4174 at 200 nM for 20 minutes, to serve as a positive control. Supplied in SDS Sample Buffer.
APPLICATIONS

Application Methods: Western Blotting

Background: Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs, such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines (1-3), and research investigators consider it an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family, as well as Mos and Tpl2/COT. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors, such as U0126 and PD98059.

$115
100 µl
The extract is prepared from whole heart tissue of adult mice, and is intended for use as a positive control in western blotting applications. The protein concentration is 2 mg/ml.The extract was prepared from whole tissue by homogenization in 1X RIPA buffer (#9806, 10X) (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 1 mM sodium EDTA, 1 mM EGTA, 1 μg/ml leupeptin, 1 mM β-glycerophosphate, 1 mM sodium orthovanadate, 2.5 mM sodium pyrophosphate), 1 mM PMSF (#8553, 34.84 mg), 1X Protease/Phosphatase Inhibitor Cocktail (#5872, 100X).The extract was subsequently sonicated and insoluble cell debris removed by centrifugation. The extract was then boiled for 5 min in 1X SDS sample buffer + DTT (#7722, Blue Loading Buffer Pack) (62.5 mM Tris-HCl pH 6.8, 10% glycerol, 2% SDS, 0.01% bromophenol blue, 42 mM DTT) to denature the proteins.
APPLICATIONS

Application Methods: Western Blotting

$64
15 ml
Cell Lysis Buffer is used to lyse cells under nondenaturing conditions.
$408
300 assays
Alexa Fluor® 488 Phalloidin allows researchers to fluorescently stain the cytoskeleton through the binding of phalloidin to F-actin. This product is intended for use on fixed and permeabilized samples due to the toxicity associated with phalloidin. After reconstitution the stock solution provides enough material to perform 300 assays based on a 1:20 dilution and a 100 μl assay volume.Alexa Fluor® 488 Fluorescent Properties: Excitation: 495, Emission: 518.
APPLICATIONS
REACTIVITY
All Species Expected

Application Methods: Immunofluorescence (Immunocytochemistry)

$408
300 assays
Alexa Fluor® 555 Phalloidin allows researchers to fluorescently stain the cytoskeleton through the binding of phalloidin to F-actin. This product is intended for use on fixed and permeabilized samples due to the toxicity associated with phalloidin. After reconstitution the stock solution provides enough material to perform 300 assays based on a 1:20 dilution and a 100 μl assay volume.Alexa Fluor® 555 Fluorescent Properties: Excitation: 555, Emission: 565.
APPLICATIONS
REACTIVITY
All Species Expected

Application Methods: Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry)

$408
300 assays
Alexa Fluor® 647 Phalloidin allows researchers to fluorescently stain the cytoskeleton through the binding of phalloidin to F-actin. This product is intended for use on fixed and permeabilized samples due to the toxicity associated with phalloidin. After reconstitution the stock solution provides enough material to perform 300 assays based on a 1:20 dilution and a 100 μl assay volume.Alexa Fluor® 647 Fluorescent Properties: Excitation: 650, Emission: 668.
APPLICATIONS
REACTIVITY
All Species Expected

Application Methods: Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry)

$61
1 mg
DAPI is supplied as a lyophilized powder in 1 mg units. It can be used to examine cellular DNA in fluorescent microscopy and cytometry applications.
APPLICATIONS
REACTIVITY
Human

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry)

$208
50 µl
$430
200 µl
APPLICATIONS

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry)

Background: DRAQ5®, 1,5-bis{[2-(di-methylamino) ethyl]amino}-4, 8-dihydroxyanthracene-9,10-dione, is a cell permeable far-red fluorescent DNA dye that can be used in live or fixed cells. This dye can be used in combination with GFP or FITC labels. DRAQ5® has been used to examine cellular DNA in flow cytometry and fluorescent microscopy applications (1-3).

$314
100 µg
ER-Tracker™ Green (BODIPY® FL Glibenclamide) is recommended for live cell imaging only; fixation with aldehydes or alcohols will inhibit staining. Excitation: 504 nm, Emission: 511 nm, Molecular Weight: 783.10 g/mol
APPLICATIONS

Application Methods: Immunofluorescence (Immunocytochemistry)