What is anti HA antibody?
Anti-HA is produced in rabbit using a synthetic peptide corresponding to amino acid residues of the human Influenza hemagglutinin (HA), conjugated to KLH. The antibody is affinity-purified on the immobilized immunizing peptide. Anti-HA antibody is specific for N- or C-terminal HA-tagged fusion proteins.
What is an HA tag used for?
The HA (hemagglutinin) tag is derived from the human influenza virus HA protein. It is well characterized and is used extensively as a general antibody epitope tag. HA tag antibodies provide a dependable method for the detection and purification of tagged target proteins without a protein-specific antibody or probe.
What is the molecular weight of HA tag?
1.1 kDa
The HA tag (YPYDVPDYA-tag) itself has a molecular weight of 1.1 kDa and is a linear epitope derived from amino acids 98-106 of the HA protein.
How do you purify HA tagged protein?
How do you purify HA-tagged proteins? HA-tagged recombinant protein can be affinity purified directly from a cell culture lysate or supernatant. The HA-tagged protein binds to the HA-tag specific monoclonal antibody conjugated on an agarose gel.
Why is SDS used in Western blotting?
SDS is generally used as a buffer (as well as in the gel) in order to give all proteins present a uniform negative charge, since proteins can be positively, negatively, or neutrally charged.
Can Ha tags be internal?
THE HA tag antibody can detect HA tags in internal, C-terminal, or N-terminal recombinant proteins. It is derived from HA molecule corresponding to amino acids 98-106.
How many kDa is HA tag?
The HA tag (YPYDVPDYA-tag) itself has a molecular weight of 1.1 kDa and is a linear epitope derived from amino acids 98-106 of the HA protein. It is used extensively as a general epitope tag in expression vectors (2, 3).
How many kDa is an HA tag?
How big is a HA tag?
The hemagglutinin (HA) tag is a 9-amino acid long peptide corresponding to residues 98-106 of the human influenza HA molecule, an ~63kDa surface glycoprotein required for the infectivity of the human virus.
Why is SDS added to transfer buffer?
Adding SDS (up to 0.1%) to the transfer buffer increases the transfer efficiency of proteins, but reduces the amount of binding to the membrane. Therefore, if SDS is added to the transfer buffer, it is important to also include methanol (10–20%).
What does SDS do to proteins?
SDS is an amphipathic surfactant. It denatures proteins by binding to the protein chain with its hydrocarbon tail, exposing normally buried regions and coating the protein chain with surfactant molecules. The polar head group of SDS adds an additional benefit to the use of this denaturant.
How do you elute a HA tag?
Use 4 X 1 column volumes of 1 mg/ml HA peptide solution to elute HA-tagged proteins. Elution yields will increase by incubating the peptide on the column for 10-15 minutes at 30-37ºC for each elution fraction. Reconstitute to 1 mg/ml using TBS or distilled water.
