Synonyms: affinity matrix, anti-ha
Storage: 2-8°C
UNSPSC Code: 12352200
RIDADR: NONH for all modes of transport
Analysis Note:
Yield of 9 nmol purified protein/ml affinity matrix was determined using a whole-cell bacterial extract containing expressed HA-tagged bacterial alkaline phosphatase.
Features and Benefits:
• Achieve enhanced specificity and reduced cross-reactivity of Anti-HA High Affinity.
• Use nondenaturing conditions for elution of your HA-tagged protein of choice.
• Purify even rarely expressed HA-tagged proteins.
• Highly specific to YPYDVPDYA.
• Suitable for purification of proteins containing HA-epitop as N-terminal, C-terminal or internal fusion.
• Applicable with crued cell extracts from mammalian, bacterial and yeast expression systems.
Application:
Use Anti-HA Affinity Matrix for:
• Immunoprecipitation of HA-tagged proteins from mammalian, bacterial, and yeast cell extracts
• Affinity column purification of HA-tagged proteins from crude protein extracts
General description:
Anti-HA Affinity Matrix is an Anti-HA monoclonal rat antibody (clone 3F10) that is immobilized. Anti-HA 3F10 recognizes the 9-amino acid sequence (YPYDVPDYA), derived from the human influenza hemagglutinin (HA) protein. The antibody recognizes its antigenic determinant even when the HA peptide epitope is introduced into unrelated recombinant proteins by a technique known as "epitope tagging".
Anti-HA Affinity Matrix: Monoclonal rat antibody (clone 3F10) is covalently coupled to agarose beads. In the coupling reaction 3.5 mg of antibody is reacted per 1 ml of beads.
Yield: Yield of purified protein is dependent upon individual expression level within crude extract. Typical yield from a 1 ml column range from 2 to 8 nmol of purified protein. Using a whole-cell bacterial extract containing HA-tagged bacterial alkaline phosphatase, column capacity was determined to be approximately 9 nmol protein/ml affinity matrix. Quantity of tagged-protein increases when additional total protein is loaded, however, a decrease in purity occur.
Reuse: Poured columns containing the affinity matrix may be used at least 10 times. Column purification using the suggested procedures with peptide elution followed by glycine stripping, was performed ten times with a bacterial extract containing HA-tagged bacterial alkaline phosphatase. No loss in column activity was observed. Activity of the column for more than ten uses should be determined by the user and depends upon the tagged protein or cell extract.
Other Notes:
For life science research only. Not for use in diagnostic procedures.
Quality:
Each lot of Anti-HA Affinity Matrix is tested for its ability to purify a HA-tagged protein expressed in stably transfected cells from a crude mammalian extract. The immunoprecipitate is analyzed by western blot using Anti-HA, High Affinity antibody.
Immunogen:
Amino acids 98-106 from the human influenza virus hemagglutinin protein
Physical form:
1 ml settled resin of Anti-HA Affinity Matrix in PBS containing 0.09% sodium azide (w/v); 2 ml suspension equals to 1 ml bed volume. One plastic column with top and bottom caps is included.
Specificity:
Anti-HA High Affinity antibody (clone 3F10) recognizes the 9-amino acid sequence YPYDVPDYA, derived from the human influenza hemagglutinin (HA) protein. This epitope is also recognized in fusion proteins that are derived from mammalian, bacterial, and yeast expression vectors, regardless of its position (N-terminal, C-terminal or internal).