The nonsignificantly differentially expressed genes are shown as black points. The significantly differentially expressed genes are plotted in red with upregulated genes on the right side and downregulated genes on the left side. Each point represents one gene, which had detectable expression in both groups (five untreated patients with HIV-1 infection versus five healthy donors). The log 2 fold change difference is presented on the x axis, and the negative log of the false-discovery rate (FDR) is presented on the y axis. Volcano plot of genes differentially expressed in PBMCs isolated from five independent healthy donors and five independent untreated patients with HIV-1 infection. (A) OTOF was induced in the PBMCs of untreated patients with HIV infection. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).įIG 1 IFN-alpha treatment induces OTOF in macrophages. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). A 37 kDa band corresponding to GAPDH was observed across the cell lines tested. The blot was probed with Anti-GAPDH Polyclonal Antibody (Product # PA1-987, 1:1,000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Note : For murine tissue samples, conjugated mouse antibody HRP-60004 and rabbit antibody 10494-1-AP are preferable.Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), COS-7 (Lane 2), MDCK (Lane 3), PC-3 (Lane 4) and tissue extract (30 µg lysate) of Ms Brain (Lane 5). GAPDH interacts with other proteins to regulate nuclear translocation and GAPDH catalytic activity. Some of What are the known protein-protein interactions of GAPDH? Useful as a control for Western blots and RT-PCR. Yes! Because GAPDH is often highly expressed throughout different tissues and cell types, GAPDH expression is Is there a reason to use an antibody targeting GAPDH if my research is not concerned with glycolysis or nuclear GAPDH is expressed primarily in the cytosolic and membrane regions, with minimal expression in the nucleus Is a key factor in regulating apoptosis, translocating to the nucleus under certain stressful conditions and actingĪs a signaling factor in oxidative stress-induced apoptosis. Other studies indicate that GAPDH plays a role in GAPDH is responsible for catalyzing the phosphorylation of glyceraldehyde-3-phosphate into D-glycerate-1,3-biphosphate. While this function of GAPDH is considered its primary function, GAPDH is also involved, evenĬritical, in other cell functions, some of which are still being studied. Recent studies have suggested that GAPDH It is recognized as an important enzyme involved in metabolic pathways, aiding in the glycolytic GAPDH stands for glyceraldehyde-3-phosphate dehydrogenase and is often referred to as a "housekeeping"
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