Signal intensity was normalized to the tubulin signal. Normalized signal intensity of GAPDH from whole cell lysates prepared from HepG2 separately transfected with two unique siRNA duplexes against GAPDH (1 & 2) or a scrambled siRNA (-). Western blot analysis of whole cell lysates prepared from HepG2 separately transfected with two unique siRNA duplexes against GAPDH (1 & 2) or a scrambled siRNA (-) probed with Rabbit anti GAPDH antibody ( VPA00187) followed by detection with HRP conjugated Goat anti Rabbit IgG (1/10,000, 1705046) and hFAB rhodamine anti-tubulin primary antibody ( 12004166) and visualized on the ChemiDoc MP with a 300 second (chemiluminescent channel) and 60 second exposure. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000. Arrow points to GAPDH (molecular weight 37 kDa). Western blot analysis of extracts of various cell lines, using GAPDH antibody (A10868). HRP conjugated Tidyblot ( STAR209P) was used at 1/200 and visualized on the Bio-Rad Chemidoc Touch Imaging System. Abpromise Western blot - Anti-GAPDH antibody - Loading Control (ab9485) Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH. Values are mean standard error of the mean. Rabbit anti GAPDH antibody ( VPA00187) was used at 1/1000 in lanes 1-5. Western blots were normalised to GAPDH and densitometric analysis was performed using image processing software. GAPDH is highly conserved across species.
Gapdh western blot plus#
8.75 μl Jurkat whole cell lysate (WCL) was run in lanes 5 & 11 while Precision Plus Protein Prestained Standards were run in lane 6. GAPDH is a housekeeping gene and used as controls in both Western blot and qPCR. IP was performed on Jurkat cell lysates using 10 µg (lanes 1 & 7), 5 µg (lanes 2 & 8), 1 µg (lanes 3 & 9) Mouse anti GAPDH antibody (MCA4739), and 10 μg Mouse IgG1 Negative Control (lanes 4 & 10) ( MCA1209).Ĩ.75 μl of each IP was loaded onto an AnykD Criterion TGX Stain-Free gel. Western blot analysis of GAPDH IP samples. Western blot: 20 g of HeLa, Mouse brain and Rat brain lysates stained with ARG65681 anti-Histone H3 antibody at 1:2000 dilution. PVDF Membrane was probed with 1 g/mL of Goat Anti-Human GAPDH-2 Antigen Affinity-purified Polyclonal. Arrow points to GAPDH (molecular weight 37 kDa). Western blot shows lysates of human testis tissue. blot analysis of GAPDH expression, detectedwith GAPDH(G-9) Alexa Fluor. Western blot analysis of whole cell lysates probed with GAPDH antibody ( VPA00187) followed by detection with HRP conjugated Goat anti Rabbit IgG (1/10,000, STAR208P) and visualized on the ChemiDoc MP with 2 second exposure. Direct near-infrared western blot analysis of -Actin expression in HeLa (A).
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