Western blot, also called an immunoblot, is an analytical assay commonly used to determine the presence of a particular protein and quantify it. The process uses polyacrylamide gel electrophoresis (PAGE) to separate proteins from a heterogenous lysate based on their molecular weight or 3D structure. The separated proteins are then transferred to a PVDF or nitrocellulose membrane, where the protein(s) of interest can be detected using highly specific antibodies. The detection can then be visualized using chromogenic, fluorescent, or chemiluminescent reporters. Since its initial discovery in the late 1970’s, this sensitive visual assay has become an invaluable tool for molecular biology and proteomics researchers.
Novus offers a range of support options for western blot when you buy a product to help you with your western blot experiment including:
![Western Blot: Human Brain Whole Tissue Lysate (Adult Normal) [NB820-59177] - Beta Actin expression on human brain whole tissue lysate using anti-Beta Actin antibody (Cat. # NBP1-47423). Image from verified customer review.](http://images.novusbio.com/fullsize/Human-Brain-Whole-Tissue-Lysate-Adult-Normal-Western-Blot-NB820-59177-img0001.jpg "Western Blot: Human Brain Whole Tissue Lysate (Adult Normal) [NB820-59177] - Beta Actin expression on human brain whole tissue lysate using anti-Beta Actin antibody (Cat. # NBP1-47423). Image from verified customer review.")
![Western Blot: Neuro2a Chloroquine Treated / Untreated Cell Lysate [NBP2-49688] - Mouse Neuoblast cells (Neuro2A) were treated with (+) and without (-) 50 uM Chloroquine overnight. Whole cell protein lysates were prepared in 1x Laemmli sample buffer and approximately 10 ug of each lysate (NBP2-49688) was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1 ug/ml anti-LC3 (NB100-2220) and 1 ug/ml anti-alpha tubulin (NB100-690) as a loading control, and detected with the appropriate secondary antibodies using chemiluminescence.](http://images.novusbio.com/fullsize/Neuro2a-Chloroquine-Treated---Untreated-Cell-Lysate-Western-Blot-NBP2-49688-img0005.jpg "Western Blot: Neuro2a Chloroquine Treated / Untreated Cell Lysate [NBP2-49688] - Mouse Neuoblast cells (Neuro2A) were treated with (+) and without (-) 50 uM Chloroquine overnight. Whole cell protein lysates were prepared in 1x Laemmli sample buffer and approximately 10 ug of each lysate (NBP2-49688) was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1 ug/ml anti-LC3 (NB100-2220) and 1 ug/ml anti-alpha tubulin (NB100-690) as a loading control, and detected with the appropriate secondary antibodies using chemiluminescence.")
![<b>Biological Strategies Validation. </b>Western Blot: Neuro2a Chloroquine Treated / Untreated Cell Lysate [NBP2-49688] - Western Blot: [NBP2-49688] - 10 ug of Chloroquine treated (+) and untreated (-) Neuro2A lysates in 1x Laemmli sample buffer (NBP2-49688) were separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1 ug/ml monoclonal anti-LC3 (NBP2--46892) and detected with an anti-rabbit secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/Neuro2a-Chloroquine-Treated---Untreated-Cell-Lysate-Western-Blot-NBP2-49688-img0004.jpg)
![Western Blot: Mouse Brain Whole Tissue Lysate (Adult Normal) [NB820-59657] - Western blot analysis of GAPDH (NB300-323) using whole normal mouse brain lysate [NB820-59657].](http://images.novusbio.com/fullsize/Mouse-Brain-Whole-Tissue-Lysate-Adult-Normal-Western-Blot-NB820-59657-img0005.jpg "Western Blot: Mouse Brain Whole Tissue Lysate (Adult Normal) [NB820-59657] - Western blot analysis of GAPDH (NB300-323) using whole normal mouse brain lysate [NB820-59657].")
![SDS-Page: Mouse Brain Whole Tissue Lysate (Adult Normal) [NB820-59657] - Ponceau S staining of whole normal mouse brain lysate [NB820-59657].](http://images.novusbio.com/fullsize/Mouse-Brain-Whole-Tissue-Lysate-Adult-Normal-SDS-Page-NB820-59657-img0004.jpg)
![SDS-Page: Mouse Brain Whole Tissue Lysate (Whole Normal) [NB820-59657] - Protein lysate is positively confirmed by Western Blot using specific biomarker: GAPDH for total protein, cytoplasmic and universal total protein; Histone for nuclear protein; Na/K ATPase for membrane protein; Vimentin for cytoskeleton protein.](http://images.novusbio.com/fullsize/Mouse-Brain-Whole-Tissue-Lysate-Whole-Normal-SDS-Page-NB820-59657-img0006.jpg)