The largest database of trusted experimental protocols

Anti rabbit or anti mouse igg horseradish peroxidase

Manufactured by Santa Cruz Biotechnology
Sourced in United States

Anti-rabbit or anti-mouse IgG–horseradish peroxidase is a conjugate used as a detection agent in various immunoassays and immunohistochemical techniques. It consists of horseradish peroxidase enzyme coupled to either anti-rabbit or anti-mouse immunoglobulin G (IgG) antibodies.

Automatically generated - may contain errors

3 protocols using anti rabbit or anti mouse igg horseradish peroxidase

1

Western Blot Protein Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
Proteins from cell lysates (50 μg protein) were separated via SDS–PAGE and electroblotted onto a polyvinylidene difluoride (PVDF) membrane for 90 min at 100 V at 4 °C. After blocking with 5% bovine serum albumin (BSA) for 60 min in TBS-T (20 mM Tris–HCl, pH 7.5, 50 mM NaCl, 0.1% Tween 20), the membrane was incubated overnight with primary antibodies in 5% BSA at 4 °C, washed with TBS-T buffer, and incubated for 60 min in anti-rabbit or anti-mouse IgG–horseradish peroxidase (Santa Cruz Biotechnology, Santa Cruz, CA, USA) secondary antibodies diluted in 5% BSA (1:3000). After treating with chemiluminescence reagents (Thermo Fisher Scientific, Rockford, IL, USA), protein bands were detected and quantified using a Kodak Gel Logic 440 imaging system with Image J software. Unless specified otherwise, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control for all western blotting analyses.
+ Open protocol
+ Expand
2

Western Blot Analysis of Protein Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
Proteins from cell lysates (50 μg protein) were separated via SDS–PAGE and electroblotted onto a polyvinylidene difluoride (PVDF) membrane for 90 min at 100 V at 4 °C. After blocking with 5 % BSA for 60 min in TBS-T (20 mM Tris–HCl, pH 7.5, 50 mM NaCl, 0.1 % Tween 20), the membrane was incubated overnight with primary antibodies in 5 % BSA at 4 °C, washed with TBS-T buffer, and incubated for 60 min in anti-rabbit or anti-mouse IgG–horseradish peroxidase (Santa Cruz Biotechnology, Santa Cruz, CA, USA) secondary antibodies diluted in 5 % BSA (1:3000). After treating with chemiluminescence reagents (Thermo Fisher Scientific, Rockford, IL, USA), protein bands were detected and quantified using a Kodak Gel Logic 440 imaging system with ImageJ software. Unless specified otherwise, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control for all western blotting analyses.
+ Open protocol
+ Expand
3

Western Blot Analysis of Neural Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
For Western blot analysis, 1.25×106 NSCs were cultured in 2.5 mL media unless specified otherwise. Proteins in cell lysates (20 μg protein) were separated via sodium dodecyl sulfate–polyacrylamide gel electrophoresis and electroblotted onto a polyvinylidene difluoride (PVDF) membrane for 90 min at 100 V at 4 °C. The membranes were blocked with 5% BSA for 60 min in TBST (20 mM Tris–HCl, pH 7.5, 50 mM NaCl, 0.1% (vol/vol) Tween 20), and incubated overnight at 4°C with primary antibodies diluted (1:1000) in TBST. After being washed in TBST buffer, the membranes were incubated for 60 min in anti-rabbit or anti-mouse IgG–horseradish peroxidase (Santa Cruz Biotechnology, Santa Cruz, CA, USA) secondary antibodies diluted (1:2000) in TBST, and the labeled proteins were detected with chemiluminescence reagents (Thermo Fisher Scientific, Rockford, IL, USA). Experiments were repeated three times. Western blots intensities of bands were quantitated using Image J software (NIH, Bethesda, MD) as previously described (Kwon et al., 2011 (link)).
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!