Gut tissue samples were lysed in RIPA buffer (Beyotime, Nantong, China), and total protein was extracted. The protein levels of tight junction proteins (Claudin, Occludin, ZO-1/tight junction protein 1) and toll-like receptor (TLR) 2 and 4 were determined by a standard western blotting method with GAPDH as a loading control. Anti-Claudin (#13050-1-AP), anti-Occludin (#13409-1-AP), anti-ZO-1 (#21773-1-AP), anti-TLR4 (#19811-1-AP), and anti-GAPDH (10494-1-AP) primary antibodies were obtained from Proteintech (Wuhan, Hubei, China). An anti-TLR2 (#bs-1919R) primary antibody was obtained from Bioss (Bioss Biotech, Beijing, China).
Anti tlr4 19811 1 ap
Manufactured by Proteintech
Sourced in China
The Anti-TLR4 antibody (19811-1-AP) is a primary antibody used for the detection of Toll-like receptor 4 (TLR4) in various applications, such as Western blotting, immunohistochemistry, and immunofluorescence. TLR4 is a cell surface receptor that plays a crucial role in the innate immune response.
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Lab products found in correlation
N benzoyl l tyrosine ethyl ester, by Merck Group (2 mentions)
Anti myd88, by Cell Signaling Technology (2 mentions)
Ripa buffer, by Beyotime (1 mentions)
Anti gapdh 10494 1 ap, by Proteintech (1 mentions)
Anti p nf κb p65, by Cell Signaling Technology (1 mentions)
Anti nf κb p65, by Cell Signaling Technology (1 mentions)
Gapdh ab181602, by Abcam (1 mentions)
As1107, by Aspen (1 mentions)
Dextran sulfate sodium (dss), by MP Biomedicals (1 mentions)
Goat anti rabbit igg, by ZSGB-BIO (1 mentions)
Anti phospho stat3 tyr705, by Cell Signaling Technology (1 mentions)
Anti stat3 10253 2 ap, by Proteintech (1 mentions)
Sucralose, by Merck Group (1 mentions)
Azoxymethane aom, by Merck Group (1 mentions)
Na benzoyl l arginine 4 nitroanilide hydrochloride, by Merck Group (1 mentions)
Anti occludin, by Proteintech (1 mentions)
Anti gapdh, by ZSGB-BIO (1 mentions)
Anti β actin, by ZSGB-BIO (1 mentions)
Goat anti mouse igg, by ZSGB-BIO (1 mentions)
Dss 36 50 kda, by MP Biomedicals (1 mentions)
4 protocols using anti tlr4 19811 1 ap
1
Gut Tight Junction and TLR Protein Analysis
2
Western Blot Analysis of TLR4 and NF-κB
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The protein concentration was determined using the BCA protein concentration detection kit (Aspen, Wuhan, China). Equal amounts of proteins were then separated on 10% SDS-PAGE and electroblotted onto a PVDF membrane. Next, the membrane was incubated overnight at 4°C with the primary antibodies: anti-TLR4 (19811-1-AP, 1:1000; Proteintech, Rosemont, IL, USA), anti-p- NF-κB p65 (#3033, 1:500; Cell Signaling Technology, Danvers, MA, USA), anti-NF-κB p65 (#8242, 1:3000; Cell Signaling Technology), and GAPDH (ab181602, 1:10000; Abcam, Cambridge, UK). After that, the membrane was probed with a secondary antibody (AS1107, 1:10000; Aspen) at room temperature. Subsequently, the bands were developed with an enhanced chemiluminescence detection kit (Aspen ) and the band intensity was analyzed using the AlphaEaseFC software.
3
Colorectal Cancer Signaling Pathway Analysis
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N-benzoyl-L-tyrosine ethyl ester (BTEE), Na-Benzoyl-Larginine 4-nitroanilide hydrochloride (BAPNA), sucralose (≥98.0% HPLC), and Azoxymethane (AOM) were purchased from Sigma-Aldrich (St. Louis, MO, USA). 4-Nitrophenyl b-D-glucopyranoside was purchased from BBI Life Sciences. DSS (MW: 36–50 kDa) was obtained from MP Biomedical (Solon, OH, USA). The antibodies used in this study were anti-TLR4 (19811-1-AP, Proteintech), anti-VEGF (19003-1-AP, Proteintech), anti-STAT3 (10253-2-AP, Proteintech), anti-Phospho-Stat3 (Tyr705) (#9145, Cell Signaling Technology), anti-MyD88 (#4283, Cell Signaling Technology), anti-TRAF6 (YT4720, Immunoway), anti-inhibitor of NF-κB alpha (IκBα) (#4814, Cell Signaling Technology), and anti-Occludin (13409-1-AP, Proteintech), as well as anti-GAPDH, anti-β-Actin, goat anti-rabbit IgG, and goat anti-mouse IgG from ZSGB-BIO Co. Ltd. (Beijing, China). All other reagents used were of analytical grade.
4
DSS-Induced Intestinal Inflammation Assay
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UCB, N-benzoyl-L-tyrosine ethyl ester, and N-α-benzoyl-L-arginine 4-nitroanilide hydrochloride were purchased from Sigma–Aldrich (St. Louis, MO, United States). DSS (36-50 kDa) was obtained from MP Biomedical (Solon, OH, United States). Enzyme linked immunosorbent assay (ELISA) kits for D-lactate, TNF-α, IL-1β, and myeloperoxidase (MPO) were from Beijing Propbs Biotechnology (Beijing, China). The antibodies used in this study were anti-TLR4 (19811-1-AP; Proteintech, Rosemont, United States), anti-MyD88 (4283; Cell Signaling Technology, Danvers, MA, United States), anti-TRAF6 (ab33915; Abcam, Cambridge, MA, United States), anti-inhibitor of NF-κB alpha (IκBα) (4814; Cell Signaling Technology), and anti-occludin (ab167161; Abcam). Anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH), goat anti-rabbit immunoglobulin G and goat anti-mouse immunoglobulin G were purchased from ZSGB-BIO Co. Ltd. (Beijing, China). All other reagents used were of analytical grade.
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