The expression level of CALR and β-actin were assessed by the western blotting assay. Anti-CALR (Rabbit, 1:1,000, Proteintech, China) and anti-β-actin (Mouse, 1:5,000, Proteintech, China) were used as the primary antibody. HRP goat anti-mouse IgG (Mouse, 1:5,000, Proteintech, China) and HRP goat anti-rabbit IgG (Rabbit, 1:6,000, Proteintech, China) were used as the secondary antibody. ECL development was used for visualization.
Goat anti mouse igg hrp
Manufactured by Proteintech
Sourced in China, United States
Goat anti-mouse IgG-HRP is a secondary antibody that binds to mouse immunoglobulin G (IgG) and is conjugated with horseradish peroxidase (HRP). This reagent is used in various immunoassay techniques to detect and quantify mouse IgG in samples.
Automatically generated - may contain errors
Lab products found in correlation
Hrp goat anti rabbit igg, by Proteintech (32 mentions)
Anti β actin, by Proteintech (5 mentions)
β actin, by Proteintech (5 mentions)
Pvdf membrane, by Merck Group (4 mentions)
Gapdh, by Proteintech (3 mentions)
Urea, by Xilong (3 mentions)
Anti gapdh, by Proteintech (3 mentions)
Enhanced chemiluminescent reagent, by Thermo Fisher Scientific (3 mentions)
Cell lysis buffer, by Abcam (3 mentions)
Mini protean tgx gel, by Bio-Rad (3 mentions)
Mouse anti gapdh, by Proteintech (3 mentions)
Polyvinylidene fluoride (pvdf), by Merck Group (2 mentions)
Ripa buffer, by Solarbio (2 mentions)
Bovine serum albumin (bsa), by Biosharp (2 mentions)
Ripa lysis buffer, by Merck Group (2 mentions)
Gapdh 60004 1 ig, by Proteintech (2 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Keygen Biotech (2 mentions)
Ripa buffer, by CWBIO (2 mentions)
Bca protein assay kit, by Beyotime (2 mentions)
Ripa buffer, by Beyotime (2 mentions)
47 protocols using goat anti mouse igg hrp
1
Quantitative Assessment of CALR Protein
2
Xuanhuang Runtong Tablets Characterization
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Xuanhuang Runtong tablets (batch number: 20200101) were purchased from Hunan Shidai Yangguang Pharmaceutical Co., Ltd. (Yongzhou, China). The voucher specimen is stored in the Chinese medicine storage cabinet of the Institute of Chinese Materia Medica, Hunan Academy of Traditional Chinese Medicine. Loperamide hydrochloride capsules (batch number: KFJ2P0P) were purchased from Xi'an Janssen Pharmaceutical Co., Ltd. (Xi'an, China). The reference substances aloin (batch number: 110787-201808, 92.4%), neohesperidin (batch number: 111857-201804, 99.4%), naringin (batch number: 110722-201805, 91.7%), echinacoside (batch number: 111670-201907, 91.8%), and aloe-epine (batch number: 110795-201007, 98%) were purchased from the China Institute for Food and Drug Control.
Alcian Blue and periodic acid Schiff stains were purchased from Visher Corporation (Changsha, China). Rabbit antimouse AQP3 primary antibodies were purchased from ABclonal (Wuhan, China). Cx43 and TLR5 primary antibodies, HRP goat antimouse IgG, HRP goat antirabbit IgG, and coraLite 488-conjugated affiniPure goat antimouse IgG (H + L) were purchased from Proteintech (Chicago, USA). SuperECL Plus was purchased from Advansta (California, USA). The IL-17A primary antibody was purchased from Abcam (Cambridge, UK). TLR5 and IL-17A primers were purchased from Shanghai Shenggong Biological Engineering Co., Ltd. (Shanghai, China).
Alcian Blue and periodic acid Schiff stains were purchased from Visher Corporation (Changsha, China). Rabbit antimouse AQP3 primary antibodies were purchased from ABclonal (Wuhan, China). Cx43 and TLR5 primary antibodies, HRP goat antimouse IgG, HRP goat antirabbit IgG, and coraLite 488-conjugated affiniPure goat antimouse IgG (H + L) were purchased from Proteintech (Chicago, USA). SuperECL Plus was purchased from Advansta (California, USA). The IL-17A primary antibody was purchased from Abcam (Cambridge, UK). TLR5 and IL-17A primers were purchased from Shanghai Shenggong Biological Engineering Co., Ltd. (Shanghai, China).
3
Western Blot Analysis of Glioma Markers
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The western blotting assay assessed the expression level of IL‐1, TGF‐β, CHI3L1, PD‐L1, and β‐actin in the control and 1000 μg·mL−1 hyaluronidase groups of the HS683 and U251 cells. Anti‐IL‐1 (Rabbit, 1 : 1000, Proteintech, Wuhan, China), anti‐TGF‐β (Rabbit, 1 : 500, Abcam, Cambridge, UK), anti‐CHI3L1 (Rabbit, 1 : 1000, Proteintech, Wuhan, China), anti‐PD‐L1 (Rabbit, 1 : 1000, Abcam, Cambridge, UK), and anti‐β‐actin (Mouse, 1 : 5000, Proteintech, Wuhan, China) were used as the primary antibody. HRP goat anti‐mouse IgG (Mouse, 1 : 5000, Proteintech, Wuhan, China) and HRP goat anti‐rabbit IgG (Rabbit, 1 : 6000, Proteintech, Wuhan, China) were used as the secondary antibody. ECL development was used for visualization.
4
Immunoreactivity of Helicobacter pylori FVpE Protein
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The immunoreactivity of FVpE protein was identified by Western blot and ELISA. Purified FVpE or H. pylori antigens (NAP, UreA, UreB, VacA, CagA) was applied to 12% SDS-PAGE and transferred onto polyvinylidene difluoride membranes (PVDF, Millipore). The PVDF membrane was incubated with Rabbit anti-H. pylori serum (Abace biology, Beijing, China) or Mouse anti-FVpE serum. After washed four times, the membrane was incubated with HRP- Goat Anti-Rabbit IgG (Proteintech) or HRP- Goat Anti-mouse IgG (Proteintech). The positive signals were monitored using Luminescence ECL detection kit (ThermoFisher). Moreover, ELISA plates were incubated overnight at room temperature with 5 μg/mL purified FVpE protein. Serums from H. pylori-infected patients or healthy human were collected and diluted 1:100 in PBS. A HRP-conjugated goat anti-human IgG (Jackson ImmunoResearch) was used as the secondary antibody.
5
Immunoblotting Analysis of Transfected U87 Cells
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Transfected U87 cells were lysed by a mixture of RIPA buffer and PMSF at the concentration of 10:1. Equal amounts (20 µg) of different proteins were put in 10% gel for electrophoresis and then transferred to polyvinylidene fluoride membranes. After being blocked with 5% milk/Tris-buffered saline plus Tween 20, membrane was incubated with primary antibodies against IFI30 (1:1000; no.ab232915, Abcam), STAT6 (1:1000; no.ab32520, Abcam), P-STAT6 (1:1000; no. 9361T, Cell Signaling Technology), IL-6 (1:1000; no. #6708, R&D) and GAPDH (1:1000; no.10494-1-AP, Proteintech) at 4°C overnight. The second day, TBST was added to wash primary antibodies followed with HRP goat anti-mouse IgG and HRP goat anti-rabbit IgG (Proteintech; 1:5000) as secondary antibody incubation 1h. Immunoreactive bands were visualized by Tanon 5200. GAPDH antibody was defined as internal control.
6
ELISA Protocol for DDR1 and DDR2 Detection
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ELISA plates were coated with 1 μg·mL−1 (100 μL per well) of His‐DDR1 or His‐DDR2 extracellular antigen in PBS and dried overnight before blocking with PBS/2% BSA (Biosharp, Hefei, China). Antibody (0.00375 μg·mL−1–10 μg·mL−1) was added and incubated at 37 °C for 2 h. Plates were washed three times with 200 μL PBS/Tween 20 (0.05%). HRP‐goat anti‐mouse IgG (Proteintech, Wuhan, Hubei, China) was added and left at 37 °C for 40 min. Following this, a further four washes in PBS/Tween 20 (0.05%) were carried out before adding 100 μL per well of TMB substrate. The reaction was stopped with 2 mol·L−1 sulfuric acid and reading at 450 nm using an ELISA plate reader.
7
Protein Extraction and Western Blot Analysis
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Protein from cells was extracted with RIPA buffer (Solarbio, Beijing, China). The sample was loaded onto 12% SDS-PAGE gel for electrophoresis and then transferred onto a PVDF membrane. After blocking by 5% skim milk at room temperature for 2 h, the membrane was incubated with primary antibody targeting NPR1 (1:2000, Thermo Fisher Scientific, Waltham, MA, USA), ITGB4 (1:1000, Proteintech, Wuhan, Hubei, China), ICAM-1 (1:1000, Proteintech, Wuhan, Hubei, China), and GAPDH (1:10,000, Proteintech, Wuhan, Hubei, China) at 4 °C overnight. Afterwards, incubation of secondary antibody HRP goat anti-rabbit IgG (1:10,000, Proteintech, Wuhan, Hubei, China) or HRP goat anti-mouse IgG (1:10,000, Proteintech, Wuhan, Hubei, China) was carried out at room temperature for 2 h. The protein band was detected by Tanon 5200 Chemiluminscent and Fluorescent Imaging System (Tanon, Shanghai, China).
8
Knee Articular Cartilage Protein Analysis
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Knee articular cartilage tissue from the femoral condyle was homogenized in a lysate containing protease inhibitors, and proteins extracted were analyzed by SDS-PAGE (10% separation gel and 4.8% concentrated gel) electrophoresis. After blocking the 5% delipidated protein, 0.05 g sample was taken and ground in liquid nitrogen. Primary antibodies HIF-1α (ab1, 5 μg/ml, Abcam); SOX9 (67439-1-Ig, 1 : 1000, protein tech); MMP13 (ab39012, 1 : 1000, Abcam); ADAMTS5 (ab41037, 1 : 500, Abcam), ACAN (ab3778, 1 : 1000, Abcam); and actin (60008-1-Ig, 1 : 5000, protein tech) were added. After washing, secondary antibody HRP goat anti-mouse IgG (SA00001-1, 1 : 5000, protein tech) and HRP goat anti-rabbit IgG (SA00001-2, 1 : 6000, American protein tech) were added. Immunocomplexes were visualized with chemiluminescence using an ECL kit. Data given above were analyzed by quantity one, a professional gray-level analysis software.
9
Modulation of Autophagy by 3-Methyladenine and Betulinaldehyde
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3-Methyladenine (Cat No: HY-19312) and Betulinaldehyde (Cat No: HY-N0084) were purchased from MCE (Shanghai, China). The rAd-mCherry-GFP-LC3B was obtained from Servicebio (Wuhan, China). The anti-AKT (Cat No: 10176-2-AP, 1:1000), anti-AKT-phosphor-Ser473 (Cat No: 66444-1-Ig, 1:3000), anti-SQSTM1 (Cat No: 18420-1-AP, 1:2000), anti-actin (Cat No: 66009-1-Ig, 1:5000), HRP goat anti-Mouse IgG (Cat No: SA00001-1, 1:5000), HRP goat anti-rabbit IgG (Cat No: SA00001-1, 1:6000) were purchased from Proteintech (Hubei, Wuhan, China). The anti-ERK (Cat No: ab184699, 1:10000) was purchased from abcam (Cambridge, UK). Anti-phosphor-ERK (Cat No: AF1015, 1:1000) was purchased from affbiotech (Cincinnati, OH, USA). The anti-LC3 (Cat No: 12741S, 1:1000) was purchased from Cell Signaling Technology (Massachusetts, USA). Anti-phosphor-STAT3-Tyr705 (Cat No: abs130918, 1:1000) was bought from absin (China). And anti-STAT3 (Cat No: GB11176, 1:1000) was obtained from Servicebio (Wuhan, China).
10
Bcl-2, Bax, and Nrf2 Protein Expression in Spleen
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The protein Bcl-2, Bax, Nrf2 in spleen was detected by western blot analysis. Protein was extracted by homogenizing spleen tissue in ice-cold RIPA buffer (Merck Milli Wave, Darmstadt, Germany), and protein of spleen was obtained by using a BCA kit (Beyotime Institute of Biotechnology, Nantong, China), which was transferred to PVDF membranes after SDS-PAGE. The primary antibodies against Keap 1 (diluted 1:3000,Proteintech, USA), Bcl-2 (diluted 1:1000, Proteintech, USA), Bax (diluted 1:5000, Proteintech, USA) and β-actin (diluted 1:5000, Proteintech, USA) were used to incubate the membranes. After incubation, the membranes were cultured using antibodies (HRP goat anti-mouse IgG, Proteintech, 1:5000 dilution, No. SA00001-1) for 2 h. ECL Chemidoc XRS (Bio-Rad, Marnesla-Coquette, France) was used to detect the blots. Photographs of the membranes were quantitative analysis by using WB imaging system (JP-K300, Jiapeng Technology, China).
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