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Validated Antibodies for Cellular Proteomics


Rigorously tested antibodies for precise and accurate detection.


siRNA-validated antibodies for cellular proteomics, Thermo Scientific Pierce Products

Thermo Scientific Pierce Validated Intracellular Antibodies are cellular proteomics antibodies that have been thoroughly tested in siRNA-mediated knockdown experiments to ensure specific Western blot detection of the target proteins. Now you can proceed with confidence knowing that you will detect the correct protein clearly and unambiguously.

Why does siRNA validation matter?

Although it is general assumed that commercially-available primary antibodies detect the specified proteins, many of these have not been adequately validated using appropriate methods. Only gene-specific methods, such as siRNA, are capable of truly validating antibody specificity in whole cell lysates. This is because carefully-planned siRNA experiments involve specific knockdown of protein expression, resulting in the absence or decrease of the specific protein relative to controls.

Are your antibodies reliable?

Our scientists have screened hundreds of antibodies using the siRNA-Western blotting technique and have found that some competitor antibodies detect the wrong target protein. A significant number of competitor antibodies were either not specific (detected multiple proteins or produced high background) or were weak (detected no protein whatsoever using the manufacturer's recommended dilutions and standard Western blotting technique).

Performance of commercial antibodies. Results for antibodies tested by specific siRNA knock-down and Western blotting. Testing included 428 antibodies to 185 different intracellular targets.

Performance

Percent of 428
Antibodies Tested

Conclusion

No band detected

27%
Antibody has finicky or weak affinity (requires very specific optimization of Western blotting technique) or is not suitable for Western blotting.
High background, or many bands detected
27%
Antibody not sufficiently specific to enable unambiguous identification and measurement of target protein.

Band of appropriate MW detected.

46%
Tests indicate that 7% of these were the wrong protein (band was not the target), indicating that validation by siRNA knock-down is required to determine if the detected band is the correct (gene-specified) protein.

Validation data you can trust...

We thoroughly test each candidate intracellular target antibody to ensure that we offer only those that perform well. We are dedicated to providing the highest quality reagents available so you can get right to the important work of researching cellular pathways.

siRNA gene and protein knock-down experiments that demonstrate specificity of Thermo Scientific Validated Antibodies for cellular proteomics targets, Protein Research Products
Thermo Scientific Target-Validated Antibodies accurately detect specific targets. Cells were transfected with ON-TARGETplus SMARTpool siRNA Reagents (ILK2 or AKT2). Cell lysates were analyzed by Western blot, comparing detection by Supplier A and B antibodies and Thermo Scientific Target-Validated Antibodies. Lane 1. Mock transfection. Lane 2. Control pool siRNA. Lane 3. Gene-specific siRNA. Arrows indicate ILK protein band (52kDa) and AKT2 (60kDa). Specific target proteins and associated knockdowns were detected by our Validated Antibodies but not by the corresponding antibodies from Suppliers A and B.

Antibody Highlights:

  • Antibodies to more than 75 common intracellular targets
  • Validated by Dharmacon ON-TARGETplus siRNA to ensure target specificity
  • Western blot application optimized for use with SuperSignal Chemiluminescent Substrates
  • Ideal for any Western blotting experiment, including siRNA Western Blotting
  • Validated for human samples, although many have multi-species specificity
  • Sufficient for 10 Western blot experiments using chemiluminescent HRP substrate
    • Supplied as frozen liquid with preservative (e.g., glycerol) and carrier protein (e.g., BSA)
    • Most items contain 100-120µL at a concentration suitable for 10 mini blots
      (optimized for 1:1000 dilution for detection with the SuperSignal Substrates)
    • Some items contain 525µL at a concentration suitable for 10 mini blots
      (optimized for 1:200 dilution for detection with the SuperSignal Substrates)

Product links by primary cellular pathway:

Pathway

Targets (links to product pages)

Receptor Signaling CAV1    CHUK    GRB2    HRAS    IKBKB    IRS1    ITGB1    LYN    MYC    NCK1    PDPK1    PLCG1    PTK2    PRKCA    RAF1    RELA    RHOA    SOS1    SRC    STAT3    STAT6
AKT AKT1    CTNNB1 (beta Catenin)    FRAP1    GSK3A    GSK3B    ILK
MAP Kinase MAPK1/MAPK3 (ERK2/ERK1)    MAPK8    MAPK14(p38)    MAP2K3
TGF-beta SMAD2    SMAD3
Apoptosis BAD    BCL2    BCL2L1    BID    BIRC4    BIRC5    CASP3    PDPK1
Cell Cycle Regulation AURKB    BUB1B    CCNB1(Cyclin B1)    CCNE1    CDK4    CDK6    CDK9    CDKN1A    CDKN1B    CDKN2A(p14)    PLK1    PPP2CA    RB1
DNA Damage ATM    CDKN1B    CHEK2    MAPK14(p38)    MAP3K7    MDM2    PLK1    RAF1    TP53
Transcription Factors E2F1   FOXO1A   JUN    MYC    NFKB1    RELA    SP1    STAT6
Co-Factors CDK9    HDAC2    HDAC3
Loading Control Proteins Actin

See also the Alphabetical Menu of Antibodies (includes complete protein names and synonyms, antibody species and types, and product numbers).

Don't see the target you're interested in? Visit the Pierce-Antibodies website (www.pierce-antibodies.com).


Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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