HA-Tag antibody

Antigen: HA-Tag

Clonality: Polyclonal

Host: Rabbit

Reactivity: tag

Conjugate: Un-conjugated

Application: ELISA, Flow Cytometry (FACS), Fluorescence Microscopy (FM)

Catalog no.: ABIN1043870

Additional Information

Alternative name: HA tag

Immunogen: This antibody was purified from whole rabbit serum prepared by repeated immunizations with the 9-aa epitope tag peptide YPYDVPDYA (114-122) from hemagglutinin influenza conjugated to KLH using maleimide. A residue of cysteine was added to the carboxy terminal end to facilitate coupling.
Immunogen Type: Peptide

Format: Liquid

Isotype: IgG

Description: Anti-HA is optimally suited for monitoring the expression of HA-tagged fusion proteins. As such, anti-HA/HA can be used to identify fusion proteins containing the HA epitope. The antibody recognizes the HA epitope tag fused to the amino- or carboxy- termini of targeted proteins, as expressed in many commonly used expression vectors. This antibody has been tested by ELISA and western blotting against both the immunizing peptide and HA containing recombinant proteins. Although not tested, this antibody is likely functional for immunoprecipitation, immunocytochemistry, and other immunodetection techniques. Affinity purification of the polyclonal antibody results in very low background levels in assays and low cross-reactivity with other cellular proteins.

Characteristics: Epitope tags are short peptide sequences that are easily recognized by tag-specific antibodies. Due to their small size, epitope tags do not affect the biochemical properties of the tagged protein. Most often, sequences encoding the epitope tag are included with the target DNA at the time of cloning to produce fusion proteins containing the epitope tag sequence. This allows Anti epitope tag antibodies to serve as universal detection reagents for any tag containing protein produced by recombinant means. This means that anti-epitope tag antibodies are a useful alternative to generating specific antibodies to identify, immunoprecipitate or immunoaffinity purify a recombinant protein. The anti-epitope tag antibody is usually functional in a variety of

Specificity: This affinity purified Anti-HA antibody is directed against the HA motif and is useful in determining its presence in various assays. This polyclonal anti-HA tag antibody detects over-expressed proteins containing the HA epitope tag. To date, this antibody has reacted with all HA-tagged proteins tested. In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins.

Sterility: sterile filtered

Application Details

Application Notes: ELISA Dilution: 1:20,000 - 1:150,000.
Immunohistochemistry Dilution: 1:500 - 1:2,000.
Western Blot Dilution: 1:2,000 - 1:10,000.

Concentration: 0.95 mg/mL

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative: 0.01% (w/v) Sodium Azide

Storage: 4°C

Storage Comment: Store vial at 4° C prior to restoration.   For extended storage aliquot contents and freeze at -20° C or below.  Avoid cycles of freezing and thawing.  Centrifuge product if not completely clear after standing at room temperature.  This product is stable for several weeks at 4° C as an undiluted liquid.  Dilute only prior to immediate use. 

Expiry Date: Expiration date is one (1) year from date of opening.

Restrictions: For Research Use only

Images

Affinity Purified anti-HA epitope tag polyclonal antibody detects HA tagged recombinant proteins by IHC on formalin fixed paraffin embedded tissue. Arrowheads point to expression of HA tagged proteins in endothelial cells of mouse aorta. Sections of 4 µm were prepared from representative paraffin blocks. Sections were then deparaffinized and rehydrated with xylene and alcohol. Citrate buffer antigen retrieval was performed for 30 min in a boiling jar. Anti-HA was diluted in blocking buffer at 1:2,000 and reacted at 4° C overnight followed by signal detection using horseradish peroxidase with DAB as the chromogenic substrate. Tissue was counterstained with Mayer's hematoxylin.

anti-HA-Tag antibody (Image 2)

Publications

Product: Field, Nikawa, Broek et al.: "Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method." in: Molecular and cellular biology, Vol. 8, Issue 5, pp. 2159-65, 1988 (PubMed).