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Traditional Western Blot

Primary Antibodies 

The success of a Western blot experiment depends on having reliable antibodies validated for Western blot. Bio-Techne's Western blot antibody portfolio includes top-cited monoclonal and recombinant primary antibodies backed by our 100% guarantee

Featured Western Blot Primary Antibodies

Alkaline Phosphatase (AP) Conjugated Horseradish Peroxidase (HRP) Conjugated Blue Marker Antibody Epitope Tag Antibodies
Knockout (KO) Validated Loading Controls Recombinant Monoclonal  Simple Western Validated 

 

Western Blot Primary Antibodies by Species

Anti-Human  Anti-Mouse  Anti-Rat Anti-Bovine 

 

Secondary Antibodies & Detection

Bio-Techne offers enzyme- and fluorochrome-conjugated secondary antibodies for use in Western blot experiments. Although enzyme-conjugated secondary antibodies, such as HRP conjugates, are often used in conjunction with chemiluminescence detection and allow for the sensitive detection (low picogram) of antigens, fluorescence-based detection methods have improved signal stability and allow for simultaneous detection of multiple proteins. In fluorescence detection a fluorophore-conjugated antibody directly emits a detectable single, whereas in both chemiluminescent or chromogenic detection the process relies on an enzyme-conjugated antibody plus the addition of a substrate or detection reagent.

Common Conjugated Secondary Antibodies for Western Blot

AP (enzymatic) Biotin (enzymatic) HRP (enzymatic) Allophycocyanin (APC)/Cy7 
DyLight 488 DyLight 550 DyLight 650 DyLight 755
FITC Janelia Fluor 549 Janelia Fluor 646 NL493
NL557 PE/Cy5.5 PE/Cy7  

 

Protocols for Traditional Western Blot

Western Blot Controls  

Controls are vital for Western blot analysis to validate the specificity of protein bands and/or to uncover the root cause of any issues. Bio-Techne provides a range of controls to ensure Western blot success.

Control Lysates Over-expression Lysates Tissue Lysates Blocking Peptides and Proteins
Loading Controls       

 

Other Western Blotting Support Products 

In addition to antibodies and lysate controls, Bio-Techne also offers an assortment of other support products to aid Western blot success, including an HRP stabilizer, sub-cellular fractionation kits, protein ladders, and Western blot membranes.

 

Western Blot Support Products

HRP Stabilizer Ponceau S Staining solution PBS Buffer Tablets  RunBlue Prestained Molecular-Weight Marker- Tri colour
Recombinant Unstained Protein Ladder  Recombinant Western Blot Protein Standards  Sub-cellular Fractionation Kits  Western Blot Membranes 
Peptide Inhibitors Other Support Products    

 

Simple Western™ Automated Western Blotting Systems

Western blots are notoriously labor-intensive and challenging to perform. To overcome the challenges of traditional Western blots, Simple Western™ from ProteinSimple, a Bio-Techne brand, offers fully automated capillary Western blot analysis. The steps traditionally performed in Western blotting techniques like separation, immobilization, washing, and detection are automatically performed inside a benchtop capillary electrophoresis machine.

  • Simple Western offers high throughput Western blot analysis with fully quantitative results on up to 25 samples in just 3 hours, or 96 samples overnight.
  • Simple Western users enjoy industry-leading NIR/IR fluorescence detection sensitivity with Stellar™ Modules on Jess™.
  • Simple Western even replaces the laborious strip and re-probe of traditional Western blot testing with RePlex™, which removes the antibodies from the first round of probing for a second round of probing with fresh antibodies or total protein normalization.
Simple Western on Jess vs. traditional Western blot

Now You Can Perform Western Blotting in Single Cells with Milo™

Are you interested in single-cell proteomics? Milo™ is our Single-Cell Western blotting technique that measures protein expression in ~1,000 single cells in a single run. Validate your RNAseq data, analyze FACS-sorted cells, and gain a rich understanding of single-cell proteomics with Milo, the World’s first Single-Cell Western blotting technique.

Related Western Blot Resources

western blot handbook cover

Western Blot Handbook

This step-by-step Western blot guide from Novus Biologicals is a great introduction to Western blot. It provides detailed information for understanding, performing, and troubleshooting a Western blot protocol.

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Western Blot Troubleshooting Resource

A comprehensive, online resource for troubleshooting the most common problems of Western blot including no signal, multiple bands, and high background.

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Development of Validated Monoclonal Antibodies

This white paper describes our validation process, including antigen design, feasibility testing, specificity and sensitivity testing, and quality control and release decisions, for our monoclonal antibodies.

Background Information