Anti human gapdh antibody

For western blot analysis, protein samples were extracted from the cells or tissues with Protein Extraction Reagent (Pierce; Thermo Fisher Scientific, Inc.). The concentrations of proteins were determined using the BCA Quantification kit (Beyotime Institute of Biotechnology, Beijing, China) for subsequent sodium dodecyl sulfate-poly-acrylamide gel electrophoresis (SDS-PAGE). The proteins (20 µg) were separated by SDS-PAGE (10%) and transferred onto a PVDF membrane. The membrane was blocked using 5% non-fat milk at 25°C for 1 h, and then incubated with primary antibodies overnight at 4°C. The antibodies used were as follows: Anti-human GAPDH antibody (cat no. ab9485; 1:5,000, Abcam, Cambridge, UK), anti-human B-cell lymphoma (Bcl-2) antibody (cat. no. ab196495; 1:2,000, Abcam), anti-human Bcl-2-associated X protein (Bax) antibody (cat. no. ab32503; 1:2,000, Abcam), anti-human caspase-3 antibody (cat. no. ab13847; 1:2,000, Abcam), anti-human caspase-9 antibody (cat. no. ab202068, 1:2,000, Abcam), anti-human MMP-2 antibody (cat. no. ab37150; 1:2,000, Abcam), anti-human MMP-9 antibody (cat. no. ab73734; 1:2,000, Abcam,). Then, the membrane was incubated with anti-goat HRP-conjugated antibody (cat. no. AR1017; 1:5,000, Boster Systems, Inc., Pleasanton, CA, USA) at 25°C for 2 h. GAPDH was used as a loading control.

Immunofluorescence was performed as described in detail recently without modifications.³⁷ (link) In brief, mouse livers were fixed with paraformaldehyde, cryoprotected in sucrose, and frozen in OCT (Tissue-Tek). Frozen liver sections (5 μm) were permeabilized in ice-cold methanol followed by room-temperature 0.1% Triton X-100, then reacted with anti-human GAPDH antibody (Abcam; #ab215227) and 4′,6-diamidino-2-phenylindole (Invitrogen; #D1306) at 0.08 ng/mL. When indicated in the figure legends, anti-GS (Origene; #TA500700) was used. For Figure 2A, anti-HAL (Sigma Prestige Antibodies; #HPA038547), anti-ASL (Invitrogen; #PA5-22300), anti-ASS (Invitrogen; #PA5-82740), and anti-CPS1 (Abcam; #ab129076) antibodies were used as indicated. For Figures 2B and 2D, anti-cytokeratin 7 (Abcam; #ab68459) was used. After immunolabeling, the images were captured and analyzed on an LSM800-Airyscan microscope using ZEN Black software.