Immunofluorescence analysis of GAPDH was done on 70% confluent log phase U2OS 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 (A21994) 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||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Purified GAPDH from human erythrocytes, purified PC from bovine liver.|
|Storage buffer||HEPES buffered saline|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||0.5-2 ug/ml|
|Immunofluorescence (IF)||1-2 ug/ml|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||0.125 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.
|Western Blot (WB)||See 1 publications below|
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.
The ¿-opioid receptor affects epidermal homeostasis via ERK-dependent inhibition of transcription factor POU2F3.
A21994 was used in western blot to investigate the role of delta-opioid receptor in human keratinocytes
|Neumann C,Bigliardi-Qi M,Widmann C,Bigliardi PL||The Journal of investigative dermatology (135:471)||2015|