Western blot solutions

IgorE Abcam Parkinson disease research

Western blot is a commonly-used technique designed to separate and identify proteins from tissue or cell extracts by molecular weight.  Western blots can be performed on whole cell lysates to determine total protein expression, or on subcellular fractions in order to determine in which cellular location the protein is expressed (e.g. lysate, nucleus, chromatin bound, mitochondrial).

Western blots can be used to look at endogenous protein expression to help understand the biology of your target of interest, and can also be used to assess the success of genetic manipulations such as knock-downs, knock-outs, knock-ins or overexpression.

To be confident in your results, you need to be confident in your antibodies. Our portfolio includes a broad range of recombinant monoclonal antibodies validated in western blot, including with knock-outs where possible.

Loading controls

Loading controls are needed for Western blot to make sure loading across samples is even and to ensure any changes in target protein expression you see across different samples types are genuine. This is especially important when looking at negative samples such as knock-out cell lines or cells treated with siRNA to knockdown gene expression. A good loading control gives you confidence that your negative sample is a true negative. This ensures your experimental findings will be to the standard required for publication in peer-reviewed journals and your work is reproducible by other scientists.

A good loading control is expressed equally in all your samples, so housekeeping genes are often chosen. Some examples for cell lysates are beta-actin, alpha tubulin, GAPDH. More specific loading controls may also be chosen depending on your experiment e.g. nucleophosmin as a nuclear loading control, or Histone H3 for chromatin samples

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Browse loading controls and tag antibodies

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Anti-tag antibodies

Anti-tag antibodies are highly specific antibodies that provide a method to localize proteins which have been fused with a tag.

Tags can help you purify proteins, and are also used when studying a novel protein for which antibodies are not yet commonly available.  Common tags include FLAG, GST, 6xHis, myc and many more. Abcam provide a wide range of anti-tag antibodies validated in a wide range of  applications, including western blot.

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Explore our full range of synuclein reagents, including recombinant antibodies, immunoassays, and bioactive proteins.

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Recombinant multiclonal antibodies

Abcam’s recombinant multiclonal antibodies combine the advantages of monoclonal and polyclonal antibodies into a single product.  Multiclonals retain the high affinity and specificity of a monoclonal with the ability to bind to multiple epitopes found in a polyclonal.  This can be highly advantageous when researching a protein with multiple modifications which are functionally important, and enables you to track the correct combination of epitopes.  Importantly, unlike a traditional polyclonal, where batch variability is an inherent feature, recombinant multiclonals retain the batch-to-batch consistency of a monoclonal ensuring you have trouble-free, consistent results throughout your research.

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Antibodies to modified targets

The post-translational modification (PTM) of proteins is an important biological process, and over 400 different types of PTMs have been identified to date.  Many of these modifications are reversible and are responsible for inducing a conformational or functional change or can occur as part of a signal transduction pathway

Some of the more common modifications, like phosphorylation, acetylation, and methylation, are involved in gene regulation and enzyme activity.  Abnormal expression of PTMs has been implicated in the occurrence and development of various neurological and metabolic diseases like Alzheimer’s disease and diabetes, as well as many cancers.

Identifying and quantifying PTMs is critical for a fundamental understanding of cell biology and for elucidating disease mechanisms and diagnosis. Characterizing PTMs heavily relies on highly specific antibodies which are an essential component for detecting and enriching PTMs.

Our range of nearly a thousand recombinant monoclonal antibodies targeting PTMs have been characterized with various techniques, including peptide array, dot blot and ELISA, to confirm both specificity and minimal cross-reactivity with unmodified or related modifications.

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