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99253
PROTAC E3 Ligase Profiling Antibody Sampler Kit
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PROTAC E3 Ligase Profiling Antibody Sampler Kit #99253

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Simple Western™ analysis of lysates (1 mg/mL) from 3T3 cells using CRBN (D8H3S) Rabbit mAb #71810. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from various cell lines using XIAP (D2Z8W) Rabbit mAb.
Western blot analysis of extracts from various cell lines using β-TrCP (D13F10) Rabbit mAb.
Western blot analysis of extracts from various cell lines using VHL Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Note that 786-O is a VHL-null cell line, demonstrating specificity of the antibody.
Western blot analysis of extracts from various cell lines using c-IAP1 (D5G9) Rabbit mAb.
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.
Western blot analysis of extracts from various cell lines using CRBN (D8H3S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using CRBN (D8H3S) Rabbit mAb.
Western blot analysis of extracts from various cell lines using KEAP1 (D6B12) Rabbit mAb.
Western blot analysis of SJSA-1, U-2 OS, and Saos-2 cells, untreated (-) or treated with Nutlin 3a (10 μM, 24 hr; +), using MDM2 (D1V2Z) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Western blot analysis of extracts from HT-29 cells, untreated (-) or treated with mithramycin A (200 nM, overnight; +) using XIAP (D2Z8W) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from HeLa cells, mock transfected (-) or transfected with human c-IAP1 (+), using c-IAP1 (D5G9) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using CRBN (D8H3S) Rabbit mAb
Western blot analysis of extracts from OVCAR8 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® KEAP1 siRNA I #5285 (+) or SignalSilence® KEAP1 siRNA II #5289 (+), using KEAP1 (D6B12) Rabbit mAb (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The KEAP1 (D6B12) Rabbit mAb confirms silencing of KEAP1 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.
Confocal immunofluorescent analysis of U-2 OS cells, untreated (left) or treated with Nutlin 3a (10 μM, 24 hr; right), using MDM2 (D1V2Z) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
Immunoprecipitation of XIAP from MCF7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or XIAP (D2Z8W) Rabbit mAb (lane 3). Lane 1 represents 10% input. Western blot was performed using XIAP (D2Z8W) Rabbit mAb. The conformation specific secondary antibody Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was using to avoid cross reactivity with the IgG heavy chain.
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.
Immunohistochemical analysis of paraffin-embedded human adenoid cystic carcinoma of the salivary gland using CRBN (D8H3S) Rabbit mAb.
Confocal immunofluorescent analysis of OVCAR8 cells using KEAP1 (D6B12) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using XIAP (D2Z8W) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using CRBN D8H3S) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded HT-29 cells, untreated (left) or mithramycin A-treated (right), using XIAP (D2Z8W) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using CRBN (D8H3S) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using XIAP (D2Z8W) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded normal human colon using CRBN (D8H3S) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human spleen using CRBN (D8H3S) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human testis using CRBN D8H3S) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using CRBN (D8H3S) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using CRBN (D8H3S) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using CRBN (D8H3S) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human normal testis (left), prostate adenocarcinoma (middle), and colon carcinoma (right) using CRBN (D8H3S) Rabbit mAb (top) or a CRBN rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on CRBN protein. The similar staining patterns obtained with both antibodies to help to confirm the specificity of the staining.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CRBN (D8H3S) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
To Purchase # 99253T
Cat. # Size Price Inventory
99253T
1 Kit  (7 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
CRBN (D8H3S) Rabbit mAb 71810 20 µl
  • WB
  • IP
  • IHC
H M R 55 Rabbit IgG
VHL Antibody 68547 20 µl
  • WB
H 18-22 Rabbit 
c-IAP1 (D5G9) Rabbit mAb 7065 20 µl
  • WB
  • IP
H 62 Rabbit IgG
KEAP1 (D6B12) Rabbit mAb 8047 20 µl
  • WB
  • IF
H M R 60-64 Rabbit IgG
MDM2 (D1V2Z) Rabbit mAb 86934 20 µl
  • WB
  • IP
  • IF
H 90 Rabbit IgG
β-TrCP (D13F10) Rabbit mAb 4394 20 µl
  • WB
  • IP
H M R Mk 62 Rabbit IgG
XIAP (D2Z8W) Rabbit mAb 14334 20 µl
  • WB
  • IP
  • IHC
H Mk 53 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The PROTAC E3 Ligase Profiling Antibody Sampler Kit provides an economical means of detecting selected PROTAC E3 ligases. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Background

PROTAC (proteolysis-targeting chimera) is a technique that uses a class of small molecules to target specific proteins for degradation (1). The small molecules are heterobifunctional, consisting of two protein binding parts joined by a linker. One part of the molecule binds a protein of interest (POI), while the other part binds to an E3 ubiquitin ligase, bringing the E3 ligase close to the POI for ubiquitination coupled protein degradation (2). The first successful PROTAC study used the chimeric PROTAC-1 molecule to recruit the β-TrCP E3 ligase to dictate the ubiquitination and degradation of MetAP2 (3). There are over 600 E3 ligases, and only a few have been successfully developed by PROTAC to degrade target proteins (4). Among them, CRBN- and VHL-based PROTAC have been extensively explored and successfully applied for multiple disease treatments (5,6). The E3 ligased MDM2, c-IAP, and XIAP are fruitful in using PROTAC strategy for cancer treatment (7-9). PROTAC design using the KEAP1 E3 ligase has been developed for degradation of BRD3, BRD4, and Tau. KEAP1 is another promising member of the PROTAC team (10,11).

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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