Immunofluorescence analysis of GAPDH was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with GAPDH Mouse Monoclonal Antibody (437000) at 1 µg/mL and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Horse, Cat, Human, Non-human primate, Sheep, Pig, Bovine|
|Published species reactivity||Mouse, Human, Not Applicable, Rhesus monkey|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant protein derived from the C-terminus region of human GAPDH protein|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1-2 ug/ml|
|Immunofluorescence (IF)||1-2 ug/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:200|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1-2 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is an enzyme involved in glycolysis. In addition to this role, it is involved in other cellular processes ranging from membrane fusion, neuronal apoptosis to cancer. GAPDH is expressed at high levels in most tissues and is therefore useful as protein loading control in western blot analysis. Additionally, the antigen is conserved in most eukaryotes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Actin remodeling by Nck regulates endothelial lumen formation.
437000 was used in western blot to study how Nck signaling contributes to vascular formation and remodeling
|Chaki SP,Barhoumi R,Rivera GM||Molecular biology of the cell (26:3047)||2015|
Quantitative proteomics reveals oxygen-dependent changes in neuronal mitochondria affecting function and sensitivity to rotenone.
437000 was used in western blot to examine the impact of oxygen levels on mitochondrial function in neurons.
|Villeneuve L,Tiede LM,Morsey B,Fox HS||Journal of proteome research (12:4599)||2013|
Nck enables directional cell migration through the coordination of polarized membrane protrusion with adhesion dynamics.
437000 was used in western blot to elucidate the role of Nck in cytoskeletal remodeling during directional migration.
|Chaki SP,Barhoumi R,Berginski ME,Sreenivasappa H,Trache A,Gomez SM,Rivera GM||Journal of cell science (126:1637)||2013|
|Rhesus monkey||Not Cited||
Membrane fusion-mediated autophagy induction enhances morbillivirus cell-to-cell spread.
437000 was used in western blot to report that morbilliviruses rapidly induce autophagy for efficient cell-to-cell spread.
|Delpeut S,Rudd PA,Labonté P,von Messling V||Journal of virology (86:8527)||2012|
Quantitative proteomics of synaptic and nonsynaptic mitochondria: insights for synaptic mitochondrial vulnerability.
437000 was used in western blot to study the susceptibility of synaptic mitochondria to damage.
|Stauch KL,Purnell PR,Fox HS||Journal of proteome research (13:2620)||2014|
A quantitative TR-FRET plate reader immunoassay for measuring autophagy.
437000 was used in western blot to develop a TR-FRET immuno-assay for the quantitative measurement of autophagy using GFP-tagged LC3B.
|Hancock MK,Hermanson SB,Dolman NJ||Autophagy (8:1227)||2012|