Western blot analyses were used to probe for protein levels present in either brain slice homogenate or secreted into slice conditioned medium. For slice homogenates, brain slices were lysed using RIPA buffer and protease inhibitors. Lysates were incubated on ice for 10 minutes and then centrifuged for 10 minutes at 4°C. All samples were normalized to protein concentration, mixed with Laemmli loading buffer (1:4), boiled for 5 minutes, and loaded onto BioRad Mini-Protean TGX precast gels. Protein was transferred to PVDF membranes and blocked with 5% bovine serum albumin (BSA) in TBST for one hour. Anti-Neuroligin-3 (NovusBio; 1:250), Anti-phospho FAK pTyr861 (Thermo Fisher Scientific; 1:500), and anti-FAK (Cell Signaling Technologies; 1:500), or anti-ADAM10 (Abcam; 1:500) were diluted in 1% BSA/TBST and incubated with the membrane overnight. Secondary anti-rabbit conjugated to HRP (BioRad) was then added for one hour (1:1000). Proteins were visualized using Clarity ECL Western Substrate (BioRad) and quantified and analyzed using ImageJ.
Anti rabbit conjugated to hrp
Manufactured by Bio-Rad
Anti-rabbit conjugated to HRP is a laboratory reagent used to detect and quantify the presence of rabbit-derived proteins or antibodies in various biological samples. The horseradish peroxidase (HRP) enzyme is covalently attached to an anti-rabbit antibody, allowing for the specific detection and visualization of rabbit-derived targets through enzymatic color reactions.
Automatically generated - may contain errors
Lab products found in correlation
Anti fak, by Cell Signaling Technology (2 mentions)
Clarity western ecl substrate, by Bio-Rad (2 mentions)
Anti adam10, by Abcam (2 mentions)
Anti neuroligin 3, by Novus Biologicals (2 mentions)
Anti phospho fak ptyr861, by Thermo Fisher Scientific (2 mentions)
Mini protean tgx precast gel, by Bio-Rad (2 mentions)
2 protocols using anti rabbit conjugated to hrp
1
Western Blot Analysis of Brain Proteins
2
Western Blot Analysis of Brain Proteins
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Western blot analyses were used to probe for protein levels present in either brain slice homogenate or secreted into slice conditioned medium. For slice homogenates, brain slices were lysed using RIPA buffer and protease inhibitors. Lysates were incubated on ice for 10 minutes and then centrifuged for 10 minutes at 4°C. All samples were normalized to protein concentration, mixed with Laemmli loading buffer (1:4), boiled for 5 minutes, and loaded onto BioRad Mini-Protean TGX precast gels. Protein was transferred to PVDF membranes and blocked with 5% bovine serum albumin (BSA) in TBST for one hour. Anti-Neuroligin-3 (NovusBio; 1:250), Anti-phospho FAK pTyr861 (Thermo Fisher Scientific; 1:500), and anti-FAK (Cell Signaling Technologies; 1:500), or anti-ADAM10 (Abcam; 1:500) were diluted in 1% BSA/TBST and incubated with the membrane overnight. Secondary anti-rabbit conjugated to HRP (BioRad) was then added for one hour (1:1000). Proteins were visualized using Clarity ECL Western Substrate (BioRad) and quantified and analyzed using ImageJ.
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