The cells were lysed by RIPA lysis buffer (Epizyme PC101) followed by centrifuging at 12,000 × g for 5 min at 4 oC. Total protein was collected and stored in −80 oC until use. Western blotting was performed as routine. The primary antibodies used were anti-human ELF3 (ABclonal A6371), anti-human PI3K (Cell Signalling Technology 4249), anti-human P-PI3K (Cell Signalling Technology 4228), anti-human AKT (ABclonal A11030), anti-human P-AKT (Cell Signalling Technology 4060), anti-human P65 (Abcam ab16502), anti-human P-P65 (Abcam ab86299), anti-human IKKA (ABclonal A0422) and anti-human P-IKKA (ABclonal AP0546). Secondary antibody was goat anti-rabbit IgG (H + L) HRP (ABways AB0101).
Goat anti rabbit igg h l hrp
Manufactured by Abways
Goat anti-rabbit IgG (H + L) HRP is a secondary antibody that binds to rabbit primary antibodies. It is conjugated with horseradish peroxidase (HRP) enzyme, which can be used for detection in various immunoassays.
Automatically generated - may contain errors
Lab products found in correlation
Ab16502, by Abcam (1 mentions)
Rabbit anti tnfα antibody, by GeneTex (1 mentions)
Rabbit anti β tubulin antibody, by Cell Signaling Technology (1 mentions)
Rabbit anti il 1β antibody, by Abcam (1 mentions)
Chemidoc xr, by Bio-Rad (1 mentions)
Enhanced bca protein assay kit, by Beyotime (1 mentions)
Ab0102, by Abways (1 mentions)
Gapdh, by Abways (1 mentions)
Srebp1, by Santa Cruz Biotechnology (1 mentions)
Lamin b1, by Proteintech (1 mentions)
3 protocols using goat anti rabbit igg h l hrp
1
Western Blot Analysis of ELF3, PI3K, and NF-κB Pathway
2
Western Blot Analysis of Brain Proteins
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Western blot analysis was performed as previously described [38 (link)]. For details, see SI Methods . Briefly, mice were sacrificed by rapid decapitation under deep anesthesia. The brain tissues were dissected in ice-cold 1× PBS buffer, then the samples were homogenized and centrifuged. Total protein concentration was determined using an Enhanced BCA Protein Assay Kit (Beyotime). The following antibodies were used: rabbit anti-PV antibody (1:4000, Abcam), rabbit anti-Arg1 antibody (1:5000, Abcam), rabbit anti-β-tubulin antibody (1:5000, Cell Signaling Technology), rabbit anti-IL1β antibody (1:2000, Abcam), rabbit anti-IL6 antibody (1:3000, GeneTex), rabbit anti-TNFα antibody (1:3000, GeneTex), and goat anti-rabbit IgG (H + L) HRP (1:8000, Abways). Protein bands were visualized using a chemiluminescence system (ChemiDocTM XRS+, BioRad). The protein expressions were semi-quantitatively analyzed with ImageJ software.
3
Western Blot Analysis of Lipid Metabolism
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Cells and tissue were rinsed once with PBS and lysis was made in ice-cold lysis buffer (250mM NaCl, 1% Triton, 25mM Hepes pH 7.4) supplemented with protease and phosphatase inhibitors. Nucleoprotein was extracted according to the manufacturer’s instructions (Solarbio). Cell lysates or immunoprecipitates were separated by SDS-PAGE and transferred to PVDF membranes. After blocking with 5% BSA, the membrane was incubated with primary antibodies at 4°C overnight, followed by incubation with the corresponding secondary antibodies for 1 h at room temperature. The membranes were washed three times for 10 min each. The following antibodies were used: GAPDH (Abways, AB0037, 1:5000), SREBP1 (Santa Cruz, sc-365513, 1:1000), Lamin B1 (Proteintech, 66095-1-Ig, 1:10000), FAS (CST, 3180S, 1:1000), pACC (CST, 11818S, 1:1000), ACC (CST, 3676S, 1:1000), CPT1A (CST, 12252S, 1:1000), ATGL (CST, 2439S, 1:1000), pAKT (CST, 4060S, 1:1000), AKT (CST, 4691S, 1:1000), Goat Anti-Rabbit IgG (H+L) HRP (Abways, AB0101, 1:5000), Goat Anti-Mouse IgG (H+L) HRP (Abways, AB0102, 1:5000). Arbitrary densitometry units were quantified by ImageJ analysis.
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