Pretreated bacteria or purified protein were mixed with loading buffer and loaded onto SDS polyacrylamide gels. After electrophoresis, gel was used for staining with coomassie brilliant blue or transferred to polyvinylidene difluoride membrane (Roche Diagnostics) for 2 hr at 100 V using a transblotting apparatus (Bio‐Rad). The membrane was blocked overnight at 4℃ in 5% skimmed milk‐TBST and was detected by His‐tag (4C2) monoclonal antibody (Bioworld Technology) with a 1:8,000 dilution at RT for 1 hr. The primary antibody binding was incubated with a 1:20,000 dilution of horseradish peroxidase (HRP)‐conjugated goat anti‐mouse IgG secondary antibody (Proteintech, China) at RT for 1 hr and visualised with an enhanced chemiluminescence kit (CWBio).
His tag 4c2 monoclonal antibody
Manufactured by Bioworld Technology
Sourced in China
The His-tag (4C2) monoclonal antibody is a laboratory reagent used to detect and purify proteins that have been engineered to contain a histidine-tag (His-tag) sequence. The antibody specifically binds to the His-tag, allowing for the identification and isolation of the tagged protein from complex samples.
Automatically generated - may contain errors
Lab products found in correlation
Polyvinylidene difluoride membrane, by Roche (2 mentions)
Transblotting apparatus, by Bio-Rad (2 mentions)
Horseradish peroxidase hrp conjugated goat anti mouse igg secondary antibody, by Proteintech (1 mentions)
Enhanced chemiluminescence kit, by CWBIO (1 mentions)
Horseradish peroxidase hrp conjugated goat anti mouse igg, by Proteintech (1 mentions)
2 protocols using his tag 4c2 monoclonal antibody
1
SDS-PAGE and Western Blot Analysis
2
Western Blot Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Pretreated bacteria or purified protein were solubilized and loaded onto SDS polyacrylamide gels. After electrophoresis, samples were stained with Coomassie brilliant blue, or transferred (2 h at 100 V) to a polyvinylidene difluoride membrane (Roche Diagnostics, German) using a transblotting apparatus (Bio-Rad, USA). The membrane was blocked in 5 % skimmed milk-TBST at 4℃ overnight. The membrane was incubated with His-tag (4C2) monoclonal antibody (1:8,000; Bioworld Technology, China) at RT for 1 h followed by horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (1:20,000; Proteintech, China) at RT for 1 h, and visualized with enhanced chemiluminescence (CWBio, China).
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