Western Blotting (WB), ELISA, Immunoprecipitation (IP)
Sequence
HHHHHH
Specificity
This protein-A purified antibody is directed against the 6X His motif and is useful in determining its presence in various assays. This monoclonal 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.
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 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.
Sterility
Sterile filtered
Immunogen
This antibody was produced in mice by repeated immunizations with 6X His epitope tag peptide H-H-H-H-H-H conjugated to KLH using maleimide. Immunogen Type: Peptide
WB, FLISA, FM
Host: Mouse
Monoclonal
33D10-D2
DyLight 649
Application Notes
Anti-6X His is optimally suited for monitoring expression of His-tagged fusion proteins. As such, anti-6X His/6X His can be used to identify fusion proteins containing the 6X His epitope. The antibody recognizes the His tag fused either to the amino- or carboxy- termini of targeted proteins. This antibody has been tested by ELISA and western blotting against both the immunizing peptide and His-containing recombinant proteins. Although not tested, this antibody is likely functional for immunoprecipitation and immunocytochemistry.
Restrictions
For Research Use only
Format
Lyophilized
Reconstitution
Reconstitution Buffer: Restore with deionized water (or equivalent), Reconstitution Volume: 100 µL
Concentration
1.0 mg/mL
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
Preservative
Gentamicin sulfate
Handling Advice
Do NOT add Sodium Azide!
Storage
4 °C/-20 °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. Expiration date is one (1) year from date of opening.
Expiry Date
12 months
Li, Xu, Wang, Gong, Liu, Zheng, Li, Li: "Epitope mapping reveals the binding mechanism of a functional antibody cross-reactive to both human and murine programmed death 1." in: mAbs, Vol. 9, Issue 4, pp. 628-637, (2018) (PubMed).
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. Supplier produces anti-epitope tag antibodies against many common epitope tags including Myc, GST, GFP, 6X His, MBP, FLAG and HA. Supplier also produces antibodies to other tags including FITC, Rhodamine (TRITC), DNP and biotin. Synonyms: HRP anti-HIS, HIS Antibody, 6X His Tag Antibody, HHHHHH epitope tag antibody