H9c2 cells and cardiac tissue were lysed with RIPA including PMSF (Phenylmethanesulfonyl fluoride, Beyotime, Shanghai, China). The concentration of protein was checked by BCA Protein Assay Kit (Beyotime, Shanghai, China). Subsequently, the protein was separated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, MA, USA). The primary antibodies included: anti-GRP78 (1:1000, proteintech, Wuhan, China), anti-CHOP and anti-F4/80 (1:1000, Cell Signaling Technology, Danvers, MA, USA), anti-Bcl2 and anti-Bax (1:500, Santa Cruz Biotechnology, Dallas, TX, USA), anti-β actin (1:1000, BOSTER, Wuhan, China), anti-GAPDH (1:1000, Beyotime, Shanghai, China). Secondary antibodies were: HRP-Conjugated AffiniPure Goat Anti-Rabbit IgG and HRP-Conjugated AffiniPure Goat Anti-Mouse IgG (1:2000, ZSGB-BIO, Beijing, China). Immunoreactive proteins were visualized using ECL Western blot detection reagents (Cell Signaling Technology, USA).
Hrp conjugated affinipure goat anti rabbit igg
Manufactured by ZSGB-BIO
Sourced in China
HRP-Conjugated AffiniPure Goat Anti-Rabbit IgG is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and bind to rabbit primary antibodies in various immunoassay techniques.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Boster Bio (3 mentions)
Hrp conjugated affinipure goat anti mouse igg, by ZSGB-BIO (3 mentions)
Bca protein assay kit, by Beyotime (3 mentions)
Ripa buffer, by Keygen Biotech (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Anti chop, by Cell Signaling Technology (1 mentions)
Anti grp78, by Proteintech (1 mentions)
Anti bcl2 and anti bax, by Santa Cruz Biotechnology (1 mentions)
Anti f4 80, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Beyotime (1 mentions)
Anti ctr1, by Abcam (1 mentions)
Sds page, by Thermo Fisher Scientific (1 mentions)
Supersignal west pico, by Thermo Fisher Scientific (1 mentions)
Anti ctr1, by Santa Cruz Biotechnology (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Mouse anti β actin monoclonal antibody, by Boster Bio (1 mentions)
Anti akt monoclonal antibody, by Cell Signaling Technology (1 mentions)
Ecl western blotting detection regents, by GE Healthcare (1 mentions)
Anti cyclin d1 monoclonal antibody, by Santa Cruz Biotechnology (1 mentions)
Anti bax monoclonal antibody, by Santa Cruz Biotechnology (1 mentions)
4 protocols using hrp conjugated affinipure goat anti rabbit igg
1
Protein Expression Analysis in Cardiac Cells
2
Quantitative Western Blot Analysis of CTR1 Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting results were quantified using Image J software. Proteins were harvested from A549, H460, H1299 and A549/cDDP cells. Cells lysed in RIPA buffer containing PMSF (protease and phosphatase inhibitors) were quantified via BCA protein assay. Proteins separated on 10% SDS-PAGE (Invitrogen) were transferred onto PVDF membranes (PolyVinylidene Fluoride). After blocking in 5% defatted milk, membranes were incubated with primary antibodies overnight at 4°C. Membranes were washed with TBST and incubated with Horse Radish Peroxidase (HRP)-conjugated secondary antibodies for 1 h at room temperature. Primary antibodies included: anti-CTR1 (1:1000, Abcam, Cambridge, Britain), anti-β-actin (1:1000, BOSTER, Wuhan, China) and anti-β-tublin (1:1000, BOSTER, Wuhan, China). Secondary antibodies included: HRP-Conjugated AffiniPure Goat Anti-Rabbit IgG (1:2000, ZSGB-BIO, Beijing, China) and HRP-Conjugated AffiniPure Goat Anti-Mouse IgG (1:2000, ZSGB-BIO, Beijing, China). Chemiluminesence western blotting reagents (Cell Signaling Technology, Danvers, MA, USA) were used to detect immunoreactive proteins. Protein bands were measured using Eagle Eye II software.
3
Western Blot Analysis of CTR1 Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the indicated treatment, the cells were harvested and lysed by RIPA buffer (KeyGENBioTECH, Nanjing, China). The protein was extracted and its concentration was measured by BCA Protein Assay Kit (Beyotime, Shanghai, China). SDS-PAGE was used to separate 60 μg of protein. Then followed by transfer onto a PVDF membrane (Millipore Corporation, MA, USA). Membranes were blocked for 1 h at room temperature with 5% non-fat dry milk in Tris-Buffered-Saline with Tween (TBST), followed by incubation overnight at 4°C with specific antibodies. After being washed 3 times for 5 min with TBST, the membrane was then incubated with appropriate secondary antibodies for 1 h at room temperature. After extensive washing with TBST, proteins were visualized by the SuperSignal West Pico (Thermo Scientific, Wilmington, DE, USA). Antibodies used include: anti-CTR1 (1:1000, Santa Cruz Biotechnology, CA, USA), anti-β-actin (1:1000, BOSTER, Wuhan, China), HRP-Conjugated AffiniPure Goat Anti-Rabbit IgG (1:2000, ZSGB-BIO, Beijing, China), HRP-Conjugated AffiniPure Goat Anti-Mouse IgG (1:2000, ZSGB-BIO, Beijing, China).
4
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the indicated treatments, total cell extracts were obtained and lysed by RIPA buffer (KeyGENBioTECH, Nanjing, China). Protein concentrations were determined according to BCA Protein Assay Kit (Beyotime, Shanghai, China). The extracted proteins in the cell lysates were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). The primary antibodies were as follows: rabbit anti-EGFR monoclonal antibody (Cell Signaling technology, Danvers, MA, USA), anti-PTEN monoclonal antibody (Cell Signaling technology, Danvers, MA, USA), anti-p-AKT(Ser473) monoclonal antibody (Cell Signaling technology, Danvers, MA, USA), anti-AKT monoclonal antibody (Cell Signaling technology, Danvers, MA, USA), anti-cyclin D1 monoclonal antibody (Santa Cruz Biotechnology, Dallas, USA), anti-Bcl2 monoclonal antibody (Santa Cruz Biotechnology, Dallas, USA), anti-Bax monoclonal antibody (Santa Cruz Biotechnology, Dallas, USA), anti-CYP2E1 monoclonal antibody (Epitomics, CA, USA) and mouse anti-β-actin monoclonal antibody (BOSTER, Wuhan, China). Secondary antibodies include HRP-Conjugated AffiniPure Goat Anti-rabbit IgG (ZSGB-BIO, Beijing, China). Immunoreactive proteins were visualized using ECL western blotting detection regents (GE Health-care, Buckinghamshire, UK).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!