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72032
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit #72032

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CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 1
Western blot analysis of extracts from various cell lines using RBX1 (D3J5I) Rabbit mAb.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 2
Western blot analysis of extracts from COS and SK-MEL-28 cell lines using CUL4A Antibody.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 3
Western blot analysis of ubiquitin, NEDD8, ISG15 and SUMO-2/3 recombinant proteins (5 ng each), using NEDD8 (19E3) Rabbit mAb, Ubiquitin (P4D1) Mouse mAb #3936, ISG15 Antibody #2743 and SUMO-2/3 Antibody #4974.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 4
Western blot analysis of extracts from HeLa, NIH/3T3, and H-4-II-E cells, untreated (-) or treated with MG-132 #2194 (10μM, 90min; +), using Ubiquitin (E4I2J) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 5
Western blot analysis of extracts from various cell lines using DDB-1 (D4C8) Rabbit mAb.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 6
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 7
Western blot analysis of extracts from various cell lines using CRBN (D8H3S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 8
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using RBX1 (D3J5I) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 9
Immunoprecipitation of CUL4A from HT-29 extracts. Lane 1 is CUL4A Antibody, lane 2 is Normal Rabbit IgG #2729, and lane 3 is 10% input. Western blot analysis was perfomed using CUL4A Antibody. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 10
Western blot analysis of lysates from HeLa, RAW, C6 and COS cells, using NEDD8 (19E3) Rabbit mAb.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 11
Western blot analysis of various recombinant linkage-specific polyubiquitin chains, recombinant free monoubiquitin (MonoUb), and recombinant linear polyubiquitin (Ub linear) (300 ng each), using Ubiquitin (E4I2J) Rabbit mAb.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 12
Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a Myc-tagged cDNA expression construct encoding full-length human DDB-1 (+), using DDB-1 (D4C8) Rabbit mAb.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 13
Immunohistochemical analysis of paraffin-embedded mouse prostate using RBX1 (D3J5I) Rabbit mAb.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 14
Immunoprecipitation of CRBN protein from TF-1 extracts. Lane 1 is CRBN (D8H3S) Rabbit mAb #71810, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control#3900, and lane 3 is 10% input. Western blot analysis was performed using CRBN (D8H3S) Rabbit mAb #71810. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as the secondary antibody.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 15
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using NEDD8 (19E3) Rabbit mAb.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 16
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using NEDD8 (19E3) Rabbit mAb.
CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit: Image 17
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using NEDD8 (19E3) Rabbit mAb, preincubated with control peptide (left) or Nedd8 Blocking Peptide #1048 (right).
To Purchase # 72032
Cat. # Size Qty. Price
72032T
1 Kit  (6 x 20 microliters) $ 476

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
CRBN (D8H3S) Rabbit mAb 71810 20 µl
  • WB
  • IP
H M R 55 Rabbit IgG
CUL4A Antibody 2699 20 µl
  • WB
  • IP
H Mk 80, 82 Rabbit 
DDB-1 (D4C8) Rabbit mAb 6998 20 µl
  • WB
H M R Mk 127 Rabbit IgG
RBX1 (D3J5I) Rabbit mAb 11922 20 µl
  • WB
  • IP
  • IHC
H M R Mk 13 Rabbit IgG
NEDD8 (19E3) Rabbit mAb 2754 20 µl
  • WB
  • IP
  • IHC
H M R Mk 9 Rabbit IgG
Ubiquitin (E4I2J) Rabbit mAb 43124 20 µl
  • WB
H M R 9-300 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit provides an economical means of detecting the individual components of a CRL4/CRBN E3 ubiquitin ligase complex, including free and conjugated forms of both NEDD8 and ubiquitin. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the CRL4/CRBN Targeted Protein Degradation Complex Antibody Sampler Kit detects endogenous levels of its target protein. NEDD8 (19E3) Rabbit mAb detects endogenous levels of both free and conjugated NEDD8 protein. The antibody does not cross-react with other ubiquitin family members, including ubiquitin, SUMO-1, SUMO-2, SUMO-3, and ISG15. Ubiquitin (E4I2J) Rabbit mAb recognizes endogenous levels of free ubiquitin and polyubiquitinated proteins. This antibody is able to detect free ubiquitin, linear polyubiquitin (M1-linked), and homotypic polyubiquitin chains consisting of K6, K11, K27, K29, K33, K48, and K63 linkages.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro44 of human CRBN protein, Gly832 of human DDB-1 protein, Gly35 of human ubiquitin protein, the amino terminus of human NEDD8 protein, and the carboxy terminus of human RBX1 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser12 of human CUL4A. Antibodies are purified by peptide affinity chromatography.

Background

Targeted protein degradation is an experimental method of drug-based protein targeting that leverages endogenous E3 ubiquitin ligases and the ubiquitin proteasome system (UPS) to selectively degrade target proteins. It is being actively explored as a therapeutic strategy to target and degrade specific proteins that contribute to disease progression (1). This approach differs from traditional small-molecule therapeutics that seek to suppress disease proteins (e.g., kinases) by sterically blocking catalytic domains. Protein-targeting chimeras (PROTACs) are the prototypical protein "degraders”. PROTACs are bivalent chemical ligands that induce proximity between a target protein and an E3 ubiquitin ligase, resulting in ubiquitination of the target protein, and its subsequent degradation by the UPS (2,3). Cereblon (CRBN) is the substrate-recognition component of a Cullin-RING-ubiquitin ligase complex (CRL4/CRBN) that was among the first to be recognized for its therapeutic potential via targeted protein degradation (4). The CRL4/CRBN complex is comprised of CRBN, DDB-1, RBX1, and the scaffold protein CUL4A; its ligase activity is dynamically regulated via the covalent modification (neddylation) of CUL4A by NEDD8 (5). In unrelated mechanistic studies of multiple myeloma drugs, it was revealed that phthalimides (e.g., thalidomide, lenalidomide) promoted CRBN-dependent recruitment, ubiquitination, and proteasomal degradation of the immunological transcription factors Ikaros (IKZF1) and Aiolos (IKZF3) (6). The discovery that phthalimides were functioning as PROTACs, by eliciting the selective degradation of what were previously considered "undruggable" protein targets, led to a rapid acceleration and expansion of research into targeted protein degradation, with the promise of novel therapies for diseases deemed largely intractable using conventional small-molecule therapies (7-9).
  1. Bond, M.J. and Crews, C.M. (2021) RSC Chem Biol 2, 725-742.
  2. Sakamoto, K.M. et al. (2001) Proc Natl Acad Sci U S A 98, 8554-9.
  3. Sakamoto, K.M. et al. (2003) Mol Cell Proteomics 2, 1350-8.
  4. Krönke, J. et al. (2014) Science 343, 301-5.
  5. Hofmann, H. et al. (2013) J Virol 87, 11741-50.
  6. Lu, G. et al. (2014) Science 343, 305-9.
  7. Shirasaki, R. et al. (2021) Cell Rep 34, 108532.
  8. Alabi, S.B. and Crews, C.M. (2021) J Biol Chem 296, 100647.
  9. Alabi, S. et al. (2021) Nat Commun 12, 920.

Pathways & Proteins

Explore pathways + proteins related to this product.

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For Research Use Only. Not For Use In Diagnostic Procedures.
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