Anti-RAGE antibody (anti-mouse, monoclonal #sc-80652) was purchased from SantaCruz Biotechnology, (Dallas, TX) anti-TLR4 (anti-mouse, monoclonal #ab22048 anti-HMGB1 antibody (anti-rabbit, monoclonal, ab79823), anti-CGRP antibody (anti-goat, polyclonal, #ab36001) and anti-peripherin antibody (anti-rabbit, #ab4666) were purchased from Abcam (Cambridge, MA). Anti-HMGB1 neutralizing antibody (Chicken IgY, # 326052233) and control antibody (Chicken IgY #326058471) were purchased from Shino test corporation (Tokyo, Japan). Anti-phospho-p44/42 MAPK antibody (anti-rabbit, monoclonal, #4370), anti-p44/42 MAPK antibody (anti-rabbit, monoclonal, #4695), anti-phospho CREB (anti-rabbit, monoclonal, #9198), horseradish peroxidase (HRP)-conjugated IgG antibody (anti-rabbit, monoclonal, #7074), HRP-conjugated IgG antibody (anti-mouse, monoclonal, #7076), anti-mouse IgG (H + L), F(ab′)2 Fragment (Alexa Fluor® 647 Conjugate) #4410, and anti-rabbit IgG (H + L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 were purchased from Cell Signaling Technology (Danvers, MA).
Hrp conjugated igg antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The HRP-conjugated IgG antibody is a laboratory reagent that consists of an immunoglobulin G (IgG) antibody covalently linked to the enzyme horseradish peroxidase (HRP). This conjugate is commonly used in various immunoassay techniques, such as enzyme-linked immunosorbent assays (ELISAs), to detect and quantify target proteins or other analytes in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti p44 42 mapk antibody, by Cell Signaling Technology (3 mentions)
Anti rage antibody, by Santa Cruz Biotechnology (2 mentions)
Horseradish peroxidase hrp conjugated igg antibody, by Cell Signaling Technology (2 mentions)
Alexa fluor 488 conjugated igg h l f ab 2 fragment, by Cell Signaling Technology (2 mentions)
Anti cgrp antibody, by Abcam (2 mentions)
Anti phospho p44 42 mapk antibody, by Cell Signaling Technology (2 mentions)
Puromycin dihydrochloride, by Merck Group (1 mentions)
Tak 242, by Cayman Chemical (1 mentions)
Hmgb1 antibody, by GeneTex (1 mentions)
Anti tlr4 antibody, by Cell Signaling Technology (1 mentions)
Fps zm1, by Cayman Chemical (1 mentions)
Anti β actin antibody, by Abcam (1 mentions)
Chicken anti hmgb1 polyclonal antibody, by Shino-Test (1 mentions)
Enhanced chemiluminescence detection kit, by Merck Group (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Radioimmunoprecipitation assay ripa buffer, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Quantity one software, by Bio-Rad (1 mentions)
β actin, by Proteintech (1 mentions)
Skim milk, by Cell Signaling Technology (1 mentions)
7 protocols using hrp conjugated igg antibody
1
Antibody-Based Protein Detection Protocol
2
Investigating HMGB1 and RAGE Signaling
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Puromycin dihydrochloride (#P9620) was purchased from Sigma-Aldrich; Merck KGaA. TLR4 antagonist TAK-242 (#13871) and RAGE antagonist FPS-ZM1 (#11909) were purchased from Cayman Chemical Co. Chicken anti-HMGB1 polyclonal antibody (#326052233) was purchased from Shino-Test. Control shRNA plasmid-A (#sc-108060), HMG-1 shRNA plasmid (#sc-37982-SH), and anti-RAGE antibody (anti-mouse, monoclonal, sc-80652) were purchased from Santa Cruz Biotechnology, Inc. HMGB1 antibody (anti-mouse, monoclonal, GTX628834) was purchased from GeneTex, Inc. Anti-phospho-p44/42 MAPK antibody (p-ERK; anti-rabbit, monoclonal, #4370), anti-p44/42 MAPK antibody (ERK; anti-rabbit, monoclonal, #4695), horseradish peroxidase (HRP)-conjugated IgG antibody (goat anti-rabbit, monoclonal, #7074), HRP-conjugated IgG antibody (goat anti-mouse, monoclonal, #7076), and Alexa Fluor 488-conjugated IgG (H+L) F(ab′)2 fragment (goat anti-rabbit, monoclonal, #4412) were purchased from Cell Signaling Technology, Inc. Anti-TLR4 antibody (anti-rabbit, polyclonal, #ab13556), anti-β-actin antibody (anti-mouse, monoclonal, #ab49900), anti-CGRP antibody (anti-goat, polyclonal, #ab36001), and Alexa Fluor 647-conjugated IgG H&L antibody (donkey anti-goat, monoclonal, #ab150135) were purchased from Abcam.
3
Western Blot Analysis of IFITM Proteins
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Cell lysates were prepared using Radio Immunoprecipitation Assay (RIPA) buffer (Sigma-Aldrich, St. Louis, MO, USA). Equal amounts of protein (20 μg protein/lane) were electrophoresed on a 10%-SDS-polyacrylamide gel and electrophoretically transferred to a polyvinylidene difluoride membrane (Millipore, Billerica, MA, USA). After blocking with 5% bovine serum albumin in Tris-buffered saline at room temperature for 1 h, the membranes were incubated with antibodies against IFITM1 (Proteintech Group Cat# 60074-1-Ig Lot# RRID:AB_2233405), IFITM2, IFITM3 (Proteintech Group Cat# 66081-1-Ig Lot# RRID:AB_11182821), and β-actin (Proteintech, Wuhan, Hubei, China) or HTNV NP (provided by the Department of Microbiology, The Fourth Military Medical University) overnight at 4°C. The membranes were then washed and incubated with HRP-conjugated IgG antibody (Cell Signaling Technology, Danvers, MA, USA) for 1 h at room temperature. The blots were developed using an enhanced chemiluminescence detection kit (Millipore, Billerica, MA, USA) and visualized using X-ray film. The blot densities were analyzed using the Quantity One software (Bio-Rad, Hercules, CA, USA). In addition, the RIPA buffer contains 50mM Tris (pH = 7.4), 150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS. Protease inhibitor cocktail (Roche, Basel, Switzerland) was added before use.
4
Western Blot Protein Detection
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Whole cell lysates were separated on 10% SDS–PAGE gels and transferred to polyvinylidene difluoride membranes (PVDF). The membrane was blocked with 5% skim milk (Neogen Co., Lansing, MI, USA) and then incubated with a 1:1000 diluent of a primary antibody in TBS with 5% BSA. The membrane was washed 5 times in TBS-T and then incubated with 1:5000 diluent of the corresponding secondary antibodies HRP-conjugated IgG antibody (Cell Signaling Technology, Danvers, MA, USA) in TBS with 5% skim milk. Target proteins were visualized by using the ECL detection system (Amersham ECL Prime Western blotting Detection Regent, GE Healthcare, Chicago, IL, USA).
5
Characterization of Receptor Targets in Cell Signaling
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Puromycin dihydrochloride (cat. No. #sc-108071), control shRNA plasmid-A (#sc108060), MCT4 shRNA plasmid (h2) (#sc-45892-SH) and anti-MCT4 antibody (anti-rabbit, monoclonal, #sc-50329) were purchased from Santa Cruz Biotechnology (Dallas, TX). Anti-phospho-p44/42 MAPK antibody (p-ERK; anti-rabbit, monoclonal, #4370), anti-p44/42 MAPK antibody (ERK; anti-rabbit, monoclonal, #4695), horseradish peroxidase (HRP)-conjugated IgG antibody (goat anti-rabbit, monoclonal, #7074), HRP-conjugated IgG antibody (goat anti-mouse, monoclonal, #7076), Alexa Fluor 488-conjugated IgG (H+L) F(ab′)2 fragment (goat anti-rabbit, monoclonal, #4412), and Alexa Fluor 488-conjugated IgG (H+L) F(ab′)2 fragment (goat anti-mouse, monoclonal, #4408) were purchased from Cell Signaling Technology (Danvers, MA). Anti-VR1 antibody (TRPV1; anti-mouse, monoclonal, #ab203103), anti-CGRP antibody (anti-goat, polyclonal, #ab36001) and Alexa Fluor 647-conjugated IgG H&L (donkey anti-goat, monoclonal, #ab150135) were purchased from Abcam (Cambridge, MA, USA).
6
Hippocampal Protein Phosphorylation Analysis
7
THP-1 cell response to Ellagic Acid treatment
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THP-1 cells were seeded in 6-well plates at 1.5 × 106 cells/mL. The cells were pretreated with or without LPS (1 μg/mL) for 1 h and then treated with ES (0, 12.5, 25, and 50 μg/mL) for 1 h. After incubation, the cells were harvested by centrifugation (1500 rpm, 5 min). Proteins were extracted by RIPA lysis Kit (Beyotime) containing phenylmethanesulfonyl fluoride (PMSF). Protein concentration in the supernatant was measured using BCA protein assay kit (Takara, Japan). Equal amounts of proteins were loaded on 10% SDS-polyacrylamide gel and transferred onto nitrocellulose membranes (Amersham Biosciences, UK). After blocking in Tris-buffered saline with 0.05% Tween 20 (TBST) containing 5% nonfat milk for 1.5 h, the membranes were incubated with primary antibodies (Cell Signaling Technology) at 4°C overnight. After washing, the membranes were incubated with HRP-conjugated IgG antibodies (Cell Signaling Technology) for 1 h. Membranes were developed using ECL Western blotting detection reagent (Bio-Rad). The band intensity was measured using the FluorChem 8000 system.
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