His 6x Epitope TAG Sequence: C-H-H-H-H-H-H antibody

Antigen: His 6x Epitope TAG Sequence: C-H-H-H-H-H-H

Clonality: Polyclonal

Host: Rabbit

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN100492

Additional Information

Immunogen: This antibody was purified from whole rabbit serum prepared by repeated immunizations with 6X His epitope tag peptide H-H-H-H-H-H conjugated to KLH using maleimide.

Format: Liquid (sterile filtered)

Description: 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 tagged protein's biochemical properties. Most often sequences encoding the epitope tag are included with 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 antibody-dependent experimental procedures. Expression vectors producing epitope tag fusion proteins are available for a variety of host expression systems including bacteria, yeast, insect and mammalian cells. Antibodies-Online Immunochemicals produces anti-epitope tag antibodies against many common epitope tags including Myc, GST, GFP, 6X His, MBP, FLAG and HA. Antibodies-Online Immunochemicals also produces antibodies to other tags including FITC, Rhodamine (TRITC), DNP and biotin.

Specificity: This affinity purified antibody is directed against 6X His motif and is useful in determining its presence in various assays. This polyclonal anti-6X His-tag antibody detects over-expressed proteins containing the 6X His epitope tag. To date this antibody has reacted with all His tagged proteins so far tested. In western blotting of bacterial extracts the antibody does not cross-react with endogenous proteins. The antibody recognizes the His-tag (His-His-His-His-His-His) fused to either the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.

Application Details

Application Notes: Recommended Dilutions: ELISA: 1:10,000 - 1:40,000 WB: 1:1,000 - 1:3,000 IHC: 1:500 - 1:2,000 OTHER: User Optimized

Concentration: 1,0 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at -20° C. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Dilute only prior to immediate use. Expiration date is one (1) year from date of restoration.

Research Area: Tags/Labels

Restrictions: For Research Use only

Images

Anti-6X His epitope tag polyclonal antibody detects His-tagged recombinant proteins by western blot. Polyclonal rabbit-anti-His epitope tag at 0.5-1.0 ug/ml was used to detect 1.0 ug of recombinant protein containing the His epitope tag. A 4-20% gradient gel was used to separate the protein by SDSPAGE. The protein was transferred to nitrocellulose using standard methods. After blocking the membrane was probed with the primary antibody for 1 h at room temperature followed by washes and reaction with a 1:2500 dilution of IRDye