PrecisionAb™ Validated Western Blotting Antibodies

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PrecisionAb™ Validated Western Blotting Antibodies

Overview

PrecisionAb™ Antibody Validation Data

Selection Criteria

PrecisionAb Antibodies are assessed in the laboratory using the most stringent validation criteria in the industry to ensure industry-leading performance and reproducibility for western blotting.

Each PrecisionAb Antibody is tested against whole-cell lysates (up to 12 cell lines) expressing proteins at endogenous levels. Western blots are run by a group of scientists who scrutinize the data based on comparison against relevant bioinformatic database references, signal-to-background ratio, and other parameters.

Only those antibodies meeting the standards for all criteria are released as PrecisionAb Antibodies.

The results are high-performing antibodies that exhibit the highest levels of specificity and sensitivity, yielding superior performance in western blotting applications. 

PrecisionAb Antibody Validation

PrecisionAb Antibody validation results. Western blot analysis of 12 whole-cell lysates probed with PrecisionAb Mouse Anti-Human PCNA Primary Antibody followed by detection with goat anti-mouse secondary antibody and visualized on the ChemiDoc™ MP Imaging System with Clarity™ Western ECL Substrate.

 
 

User Testing Protocol

PrecisionAb Antibodies were evaluated by working proteomics researchers in side-by-side competitive testing. Each test site was asked to evaluate five PrecisionAb Antibodies using targets selected from ~35 antibodies based on research interests. Each PrecisionAb Antibody was used in western blotting alongside antibodies from two other vendors, each used according to the manufacturer’s recommendations.

The researchers were asked to select the competing antibodies as they normally would, by looking at publications and/or information on vendor websites. Researchers used their vendor-recommended protocols with samples provided in the form of whole-cell lysates, all using Bio-Rad’s Criterion™ TGX™ Gels, the Trans-Blot® Turbo™ Transfer System, and nitrocellulose membrane stacks.

 
 

Selected User Testing Results

 

Prof. Aldrin V. Gomes

Results Summary: Prof. Aldrin V. Gomes
University of California, Davis

Prof. Gomes studies molecular mechanisms of signal transduction in muscle contraction and cardiovascular disease, relying heavily on biophysical proteomic techniques. As an editor of several journals, he has observed that low-quality western blot data is routinely submitted, leading many to question the accuracy of protein quantification by western blotting. He has deeply examined key success factors in western blotting, including antibody selection (Ghosh et al. 2014, Gilda and Gomes 2013, Gomes 2009, Gomes 2014).

The Gomes Lab conducted side-by-side testing of PrecisionAb Antibodies with antibodies for the same targets from two leading antibody suppliers. Prof. Gomes summarized the results as follows:

"Antibodies provided from Bio-Rad were of very high quality for 80% of the antibodies tested, suggesting most antibodies will be great for western blotting. The other antibody tested was of high quality, on par with the best antibodies available for the target protein."

Side-by-side testing of anti-human BCL-2 antibodies with Jurkat cell lysate

Side-by-side testing of anti-human Bcl-2 antibodies with Jurkat cell lysate. Left, the western blot and the stain-free total protein imaging data for each antibody. Right, the relative band intensities for each of the western blots; the PrecisionAb Antibody shows ~5x greater signal intensity even at higher dilution. Primary antibody dilutions: PrecisionAb, 1:2,000; Competitor 1, 1:1,000; Competitor 2, 1:500. Secondary antibody dilution was 1:10,000 for all blots.

 
 

Prof. Ernesto Diaz-Flores

Results Summary: Prof. Ernesto Diaz-Flores
University of California, San Francisco

Prof. Diaz-Flores and colleagues in the Department of Pediatrics study primary patient leukemia samples to identify novel genetic alterations and incorporate them into algorithms for diagnosis, residual disease monitoring, and the identification of novel therapeutic approaches, focusing on juvenile myelomonocytic leukemia (JMML) and acute lymphoblastic leukemia (ALL) (Holmfeldt et al. 2013). Their identification of Bcl-2 as a promising therapeutic target has led them to test specific classes of targeted Bcl-2 inhibitors in hypodiploid ALL cell lines and patient-derived xenografts.

"To test the role of Bcl-2 in cell survival as well as the effect of blocking Bcl-2 in such cells, we needed to compare cells expressing high levels of Bcl-2 against those with low Bcl-2 levels. To do so, we measured Bcl-2 levels in 12 human cell lines using the PrecisionAb BCL-2-specific antibody provided by Bio-Rad. We compared the PrecisionAb Antibody to our best commercially available Bcl-2 antibody, and the Bio-Rad antibody gave great performance at a lower concentration than our reference antibody.

For our biochemical experiments, we select high-performance reagents that give us the best signal-to-noise ratio. Bio-Rad antibodies against Bcl-2 are high-performance antibodies, as indicated by single, clean, and clear bands, with no noticeable background signal at a high dilution of 1:1,000."

Side-by-side testing of anti-human BCL-2 and Bax antibodies with 12 different cell lysates

Side-by-side testing of anti-human BCL-2 antibodies with 12 different cell lysates.
In parallel western blots, the PrecisionAb Bcl-2 Antibody showed equal or greater signal intensities even at a higher dilution.

 
 

References

Ghosh R et al. (2014). The necessity of and strategies for improving confidence in the accuracy of western blots. Expert Rev Proteomics 11, 549–560.

Gilda JE and Gomes AV (2013). Stain-Free total protein staining is a superior loading control to β-actin for western blots. Anal Biochem 440, 186–188.

Gomes AV (2009). The Western Blot Troubleshooting Guide. NPB Department, University of California, Davis.

Gomes AV (2014). Can We Trust Western Blots? Presentation at Experimental Biology 2014, San Diego.

Holmfeldt L et al. (2013). The genomic landscape of hypodiploid acute lymphoblastic leukemia. Nat Genet 45, 242–252.

Resolve Your Western Blotting Woes with Better Antibodies
Pass vs. Fail Antibodies
High-Throughput Antibody Validation

High-Throughput Validation of Antibodies for Cancer Research

Bio-Rad research poster presented at the American Association for Cancer Research 2016 Annual Meeting

Download PDF
 

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The PrecisionAb Antibody product line is a collection of highly specific and sensitive primary antibodies (monoclonal and polyclonal) that have been extensively validated for western blotting.

Features and Benefits of PrecisionAb Antibodies

  • Robust validation — performed on up to 12 different cell lysates with endogenous protein levels
  • Reproducible data — stringent batch-to-batch QC
  • High specificity and sensitivity — guarantee low nonspecific binding and strong signal from target proteins
  • Trial size available — assess antibody performance before buying larger quantity
  • Positive control lysate — benchmarks the molecular weight of the target protein

Product Options

Formats Minimum Number of Applications Size (µl)
Trial size antibody with control lysate Two western blots 20
Antibody only Ten western blots 100

How to Order

Validation Video

Learn about our rigorous
validation process

Download the poster and watch the video of Dr. Mark Shulewitz as he walks you through the poster which discusses the validation steps for our premium line of PrecisonAb Primary Antibodies.

Watch Video | Download Poster


User Validation

  • Prof. Aldrin V. Gomes

    Side-by-side testing of anti-human Bcl-2 antibodies with Jurkat call lysate

    “Antibodies provided from Bio-Rad were of very high quality for 80% of the antibodies tested, suggesting most antibodies will be great for western blotting. The other antibody tested was of high quality, on par with the best antibodies available for the target protein.”

    Prof. Aldrin V. Gomes, University of California, Davis

    » View validation data

  • Prof. Ernesto Diaz-Flores

    Comparing Bcl-2 levels in 12 human cell lines using the PrecisionAb BCL-2-specific antibody

    “For our biochemical experiments, we select high-performance reagents that give us the best signal-to-noise ratio. Bio-Rad antibodies against Bcl-2 are high-performance antibodies, as indicated by single, clean, and clear bands, with no noticeable background signal at a high dilution of 1:1,000.”

    Prof. Ernesto Diaz-Flores, University of California, San Francisco

    » View validation data

Citations

PrecisionAb Antibodies have helped identify key proteins on western blots in the following published work:

  1. Serban AI et al. (2016). RAGE and TGF-β1 cross-talk regulate extracellular matrix turnover and cytokine synthesis in AGEs exposed fibroblast cells. PLOS ONE. PMCID: PMC4807770
  2. Serban AI et al. (2015). AGEs-Induced IL-6 synthesis precedes RAGE up-regulation in HEK 293 cells: An alternative inflammatory mechanism? Int J Mol Sci 16, 20100–20117. PMCID: PMC4613191
 

More Information

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Technical Resources

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10048952 Instruction Protocol for PrecisionAb™ Antibodies Customers Click to download
10049129 Experimental Protocol for Validating PrecisionAb™ Antibodies Using the V3 Western Workflow™ Click to download
6830 High-Throughput Validation of Antibodies for Cancer Research Click to download
6599 Antibodies You Can Rely On Flier Click to download [ Add to Cart (Free) ]
6831 Cancer Research Antibodies You Can Rely On Flier Click to download
6832 Neuroscience Research Antibodies You Can Rely On Flier Click to download

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