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Anti glyceraldehyde 3 phosphate dehydrogenase gadph

Manufactured by Cell Signaling Technology

Anti-glyceraldehyde 3-phosphate dehydrogenase (GADPH) is a lab equipment product that detects the presence and quantifies the levels of GADPH, a crucial enzyme involved in glycolysis, the metabolic pathway that converts glucose into energy. This product can be used to measure GADPH expression in various biological samples.

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2 protocols using anti glyceraldehyde 3 phosphate dehydrogenase gadph

1

Plasmid Transfection and Immunoblot Analysis

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Plasmid transfections were performed using Lipofectamine 2000 (Invitrogen) as per the manufacturer's protocol. Relative levels of different proteins were compared by immunoblot analysis. Respective empty vectors were used to normalize transfection. After 48 h of transfection, cells were lysed in RIPA lysis buffer (Cell Signaling Technology), and protein estimation was done using the BCA Protein Estimation Kit (Pierce Biotechnology, Inc.). The primary antibodies used were anti-Myc (Clontech), anti-glyceraldehyde 3-phosphate dehydrogenase (GADPH) (Cell Signaling Technology), and anti-CD4 (Novus Biologicals). The secondary antibodies used were anti-rabbit/mouse-horse radish peroxidase conjugated (Jackson ImmunoResearch). The proteins of interest were detected with EZ western horseradish peroxidase substrate (Biological Industries, Israel). GAPDH was used as a loading control in all cases.
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2

Analyzing Vaccinia Virus Infection in BMDCs

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Bone marrow-derived dendritic cells (BMDCs) were generated according to the protocol (Dai et al., 2014). BMDCs (1 × 106) from WT and KO mice were infected with MVA at a MOI (multiplicity of infection) of 10 or an equivalent amount of Heat-MVA, or UV-MVA. Whole-cell lysates were prepared. Equal amounts of proteins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the polypeptides were transferred to a nitrocellulose membrane. Phosphorylation of IRF3, IRF3, and STING levels were determined using respective antibodies (Cell Signaling). Vaccinia E3 protein level was determined by using anti-E3 monoclonal antibody (MAb 3015B2) kindly provided by Dr. Stuart N. Isaacs (University of Pennsylvania) (43 (link)). Anti-glyceraldehyde-3-phosphate dehydrogenase (GADPH) or anti-β-actin antibodies (Cell Signaling) were used as loading controls. B16-F10 melanoma cells were infected with MVA at a MOI of 10 or with an equivalent amount of Heat-MVA. Cell lysates were collected at various times post infection. Western blot analysis was performed using anti-phospho-IRF3, anti-IRF3, and anti-GAPDH antibodies as described above.
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