Table of Contents
What is V5 antibody?
V5 antibodies are immunological tools used to detect the V5 epitope tag. Derived from a small epitope found on the P and V proteins of the paramyxovirus of the simian virus 5 family, the V5 tag can vary in size from 9 to 14 amino acids. V5 antibodies can detect recombinant proteins conjugated with a V5 tag.
What is the V5 Tag?
V5 tag is a short peptide tag for detection and purification of proteins. The V5 tag can be fused/cloned to a recombinant protein and detected in ELISA, flow cytometry, immunoprecipitation, immunofluorescence, and Western blotting with antibodies and Nanobodies.
How big is his tag?
His-tags, due to their relatively small size (∼2.5 kDa), are not believed to significantly interfere with the function and structure of a majority of proteins.
How do I remove his tag?
His-tag removal from protein using TEV Protease
- Dialyze the protein against 20 mM Tris-HCl, pH 7.5.
- Determine the protein concentration.
- Combine 15 μg of protein and H2O (if necessary) to make a 45 μl total reaction volume.
- Add 5 μl of TEV Protease Reaction Buffer (10X) to make a 50 μl total reaction volume.
Does his tag affect protein activity?
The His-tag expression systems are widely used because His-tags have a low molecular weight and do not affect protein structure and functions. This means that it is not necessary to separate the His-tag from the target protein [3, 4]. Moreover, His-tag fusion proteins can easily be purified by Ni-NTA affinity resin.
What is the advantage of fluorescence Labelling?
Fluorescent labels offer many advantages, as they are highly sensitive even at low concentrations, are stable over long periods of time, and do not interfere with the function of the target molecules. The targeted imaging of labeled cells enables tracking them in vitro and in vivo.
What are fluorescent markers?
Fluorescent markers are specific molecules, like protein, which are covalently bound fluorophores that selectively bind to a functional group of the target for detection. The most commonly used fluorescent molecules are antibodies.
What is the specificity of V5 tag antibody-ChIP grade (ab9116)?
ELISA: V5 tag antibody – ChIP Grade (ab9116) specificity was verified by ELISA against the peptide conjugated to BSA (GKPIPNPLLGLDST-BSA). A 1:50,000 dilution of the antibody gave an O.D.=1.0 in a 15 minute reaction using HRP- conjugated goat anti rabbit (IgG) at 1:10000 and TMB as the substrate. Appropriate specificity controls were run.
What is the incubation temperature for v5 tag mouse monoclonal antibody?
The cells were labeled with V5 Tag Mouse Monoclonal Antibody ( Product # 37-7500) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Rabbit Anti-Mouse IgG Secondary Antibody ( Product # A-11059) at a dil… View more Store at 4°C short term.
What is V5 antibody used for?
This antibody has also been used successfully to immunoprecipitate fusion proteins that contain theV5 epitope.
What is the V5 tag for?
The V5 tag is usually used with all 14 amino acids (GKPIPNPLLGLDST), although it has also been used with a shorter 9 amino acid sequence (IPNPLLGLD). Lanes 1-3 : Cells transfected with v5-tagged protein. Lanes 4-5 : Untransfected cells. Mouse ES cells lysate. Publishing research using ab9116?