Immunofluorescent analysis of GAPDH (red) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a DyLight 650-conjugated GAPDH monoclonal antibody (Product # MA5-15738-D650) at a dilution of 1:50 for at least 1 hour at room temperature. F-actin (green) was stained with DyLight 488 Phalloidin (Product # 21833) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Bacteria, Chicken, Hamster, Human, Insect, Mouse, Rabbit, Rat, Yeast|
|Host / Isotype||Mouse / IgG1|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:25 - 1:100|
* 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.
MA5-15738-D650 has successfully been used for immunofluorescence. DyLight 650 has an excitation/emission of 652/672nm.
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.
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