Cell lysates were harvested in lysis buffer including protease inhibitors (Roche) as described previously (Lv et al., 2017 (link)). Protein concentration was determined using the Bradford assay (Biorad), and proteins were separated in SDS 4–20% polyacrylamide gels and then transferred onto a PVDF membrane. Membranes were blocked in TBS containing 5% milk, and 0.1% Tween 20 solution. Membranes were then incubated in the following primary antibodies: mouse anti-RTA (Argene, Cat# 11–008, Discontinued), anti-β-actin (Sigma, Cat # A5441), rabbit anti-PU.1 antibody (Cell Signaling Tech, Cat # 2258), rabbit anti-IRF8 antibody (Cell Signaling Tech, Cat # 5628), rabbit anti-PIAS1 antibody (Abcam, Cat # 109388), anti-HA-HRP (Cell Signaling Tech, Cat# 14031), anti-V5-HRP antibody (Thermo Fisher, Cat # R961–25) and anti-Flag-HRP antibody (Cell Signaling Tech, Cat# 86861 and Cat # 2044). The secondary antibodies (1:5,000 dilutions) used were horseradish peroxidase (HRP)-labeled anti-rabbit antibody (Jackson ImmunoResearch, Cat # 111–035-144), HRP-labeled anti-mouse antibody (Jackson ImmunoResearch, Cat # 111–035-166), HRP-labeled anti-rabbit antibody, light chain specific (Jackson ImmunoResearch, Cat # 211–032-171).
Hrp labeled anti mouse antibody
Manufactured by Jackson ImmunoResearch
HRP-labeled anti-mouse antibody is a secondary antibody conjugated with horseradish peroxidase (HRP). It is used to detect and quantify mouse primary antibodies in various immunoassay applications, such as ELISA, Western blotting, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti pias1 antibody, by Abcam (1 mentions)
Anti v5 hrp antibody, by Thermo Fisher Scientific (1 mentions)
Anti ha hrp, by Cell Signaling Technology (1 mentions)
Hrp labeled anti rabbit antibody, by Jackson ImmunoResearch (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Protease inhibitor, by Roche (1 mentions)
Bradford assay, by Bio-Rad (1 mentions)
2 protocols using hrp labeled anti mouse antibody
1
Western Blot Analysis of Protein Targets
2
Immunohistochemical Profiling of Colorectal Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
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IHC was performed using SIRT1, CD24 antibodies (Supplementary Table S2) at the dilution of 1:200 and 1:100, respectively, on the human colorectal cancer tissue and xenograft tissue sections. Briefly, unstained tissue sections were deparaffinized and rehydrated. Antigen Retrieval Solution (sodium citrate buffer 10 mmol/L, pH 6.0) was used for antigen retrieval. Endogenous peroxidase was blocked by 3% hydrogen peroxide solution or peroxidase blocker (bio SB). Tissue sections were incubated with primary antibody overnight at 4 C. Ready-to-use solution with polydetector plus link anti-mouse/rabbit and anti-mouse/rabbit horseradish peroxidase (HRP)-labeled (Bio SB) or HRP-labeled anti-mouse antibody (1:200, Jackson ImmunoResearch) was applied at room temperature for 15-30 minutes. Peroxidase was detected by 3,3 0 -diaminobenzidine (DAB) chromogen solution (Bio SB). The IHC slides were counterstained by hematoxylin.
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