LYC, complete Freund’s adjuvant (CFA), and corn oil were obtained from Solarbio (Beijing, China). The primary antibodies, including anti-GAPDH, anti-p-PI3K, anti-PI3K, anti-p-Akt, anti-Akt, anti-p-m-TOR, anti-m-TOR, anti-Bax, anti-bcl-2, anti-E-cadherin, anti-N-cadherin antibodies (at 1:1000 dilution respectively), HRP-labelled goat anti-mouse IgGs (at 1:2000 dilution), and antibodies for immunofluorescence staining were purchased from Abcam (Cambridge, MA, USA). Dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, MO, USA).
Hrp labelled goat anti mouse iggs
Manufactured by Abcam
Sourced in United States
HRP-labelled goat anti-mouse IgGs are secondary antibodies that specifically bind to mouse immunoglobulin G (IgG) antibodies. The antibodies are conjugated with horseradish peroxidase (HRP), an enzyme that can be used for colorimetric or chemiluminescent detection in various immunoassays and immunohistochemical applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti p akt, by Abcam (2 mentions)
Anti n cadherin, by Abcam (2 mentions)
Anti bcl 2, by Abcam (2 mentions)
Anti e cadherin, by Abcam (2 mentions)
Anti bax, by Abcam (2 mentions)
Anti akt, by Abcam (2 mentions)
Corn oil, by Solarbio (1 mentions)
Anti p mtor, by Abcam (1 mentions)
Complete freund s adjuvant cfa, by Solarbio (1 mentions)
Anti p pi3k, by Abcam (1 mentions)
Anti mtor, by Abcam (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Anti pi3k, by Abcam (1 mentions)
Bio spectrum gel imaging system, by Analytik Jena (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Bradford protein assay, by Bio-Rad (1 mentions)
Anti β actin, by Abcam (1 mentions)
Ab49849, by Abcam (1 mentions)
3 protocols using hrp labelled goat anti mouse iggs
1
Molecular Mechanisms of LYC Treatment
2
Western Blot Analysis of Cell Signaling
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Cells were treated with indicated reagents for 48 h. Protein lysates were harvested using ice-cold RIPA buffer (Beyotime Institute of Biotechnology, Beijing, China) containing protease inhibitor cocktail (Roche, Switzerland). The protein concentrations were quantified by Bradford Protein Assay (Bio-Rad Laboratories, USA). Then protein samples were separated by SDS-PAGE and transferred onto a PVDF membrane. Nonspecific binding sites were blocked with 5% (w/v) fetal bovine serum at room temperature for 2 h. Then, the membrane was incubated with a mouse monoclonal antibody at 4°C overnight. The primary antibodies (1:1000; Abcam, Cambridge, MA, USA) were as follows: anti-β-actin, anti-p-Akt, anti-Akt, anti-Bax, anti-bcl-2, anti-E-cadherin and anti-N-cadherin. After three washes with TBST buffer, the membranes were incubated with HRP-labelled goat anti-mouse IgGs (1:2000; Abcam) at room temperature for 1 h. Protein expression was detected by ECL and photographed by BioSpectrum Gel Imaging System (UVP, Upland, CA). Beta-actin was used as a loading control.
3
Western Blot Analysis of AKAP12 Expression
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Protein was extracted from tissues using RIPA buffer containing 1/10 Complete Mini protease inhibitor cocktail (Roche, Basel, Switzerland). The protein content was measured with a BCA Protein Assay kit (Beyotime, Shanghai, China). After denaturing at 95°C for 10 minutes, 50 μg of each protein sample was separated on a 12% SDS‐polyacrylamide gel and then transferred to a polyvinylidenefluoride (PVDF) membrane. After blocking with 5% degrease milk in TBST buffer, the membrane was incubated with mouse monoclonal antibody to AKAP12 (ab49849, 1:200; Abcam, Cambridge, MA, USA) and GAPDH (ab8245, 1:500, Abcam) at 4°C overnight. After washing 3 times with TBST buffer every 15 minutes, the membrane was further incubated with HRP‐labelled goat antimouse IgGs (1:200; Abcam) at 37°C for 1 hour. Finally, the results of Western blot were scanned with Chemical Mp Imaging System (Bio‐Rad, Hercules, CA, USA) and analysed by Gel‐ProAnalyzer (software version 4; UnitedBio, USA). GAPDH was used as a loading control for Western blots.
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