The largest database of trusted experimental protocols

Anti gapdh 3h12

Sourced in United States

Anti-GAPDH (3H12) is a monoclonal antibody that specifically recognizes the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein. GAPDH is a widely expressed enzyme involved in the glycolytic pathway and is commonly used as a housekeeping gene for normalization in various biological assays.

Automatically generated - may contain errors

2 protocols using anti gapdh 3h12

1

Immunoblot Analysis of Protein Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in radioimmunoprecipitation assay (RIPA) buffer containing 1% NP-40, 1 mM EDTA, 50 mM Tris-HCl (pH 7.4) and 150 mM NaCl. After sonication and centrifugation of the lysates, proteins (20 µg) were subjected to SDS-PAGE in 7.5% poly-acrylamide gels, followed by immunoblot analysis. The following antibodies were used: anti-BRG1 (a gift from Dr. Tsutomu Ohta, National Cancer Center, Japan), anti-NS (A300–600A; Bethyl Laboratories, Montgomery, TX, USA), anti-GAPDH (3H12; Medical & Biological Laboratories (MBL), Nagoya, Japan), anti-CD133 (W6B3C1; Miltenyi Biotec, Bergisch Gladbach, Germany) and anti-CD44 (2C5; R&D Systems, Minneapolis, MN, USA). Signals were detected by LAS-3000 (Fujifilm, Tokyo, Japan), quantified with ImageJ software (National Institutes of Health, USA) and normalized using GAPDH loading control.
+ Open protocol
+ Expand
2

Western Blot Analysis of SARS-CoV-2 Spike Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cells were lysed using radioimmunoprecipitation assay (RIPA) buffer (Cat# 16488-34, Nacalai Tesque) containing protease inhibitors. Proteins were separated using SDS-PAGE and transferred to Immobilon-P membranes (Cat# IPVH00010, EMD Millipore, Billerica, MA, USA). After blocking with 5% milk in PBS-T for 1 h, membranes were incubated with primary antibodies at 4°C overnight. Membranes were then washed with PBS-T and incubated with secondary antibodies for 2 h. Membranes were washed again with PBS-T and immunoblot signals were developed using EzWestLumi plus (Cat# WES-7120, ATTO, Tokyo, Japan) and recorded using an ImageQuant LAS4000 mini-image analyzer (GE Healthcare Japan, Tokyo, Japan). The antibodies used were as follows: anti-spike antibody 1A9 (Cat# GTX632604, GeneTex, CA, USA), anti-cleaved spike (Ser 686) antibody (Cat# 84534, Cell Signaling Technology, Massachusetts, USA), anti-GAPDH 3H12 (Cat# 171-3, Medical & Biological Laboratories, Aichi, Japan), goat anti-mouse IgG antibody conjugated with horseradish peroxidase (Cat# SA00001-1, Proteintech Group, Rosemont, USA), donkey anti-rabbit IgG antibody conjugated with horseradish peroxidase (Cat# NA934, Cytiva, Tokyo, Japan).
+ 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!