His Tag Antibody

• Specificity
  Detects proteins containing accessible consecutive histidine regions. The antibody detects His tags localized at the amino- or carboxyl‑terminus.
• Source
  Monoclonal Mouse IgG₁ Clone # AD1.1.10
• Purification
  Protein A or G purified from hybridoma culture supernatant
• Immunogen
  His-tagged peptide
• Formulation
  Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Endotoxin Level
  <0.20 EU per 1 μg of the antibody by the LAL method.
• Label
  Unconjugated

• Western Blot
  0.2 µg/mL
  See below
• Simple Western
  10 µg/mL
  See below
• Flow Cytometry
  0.25 µg/10⁶ cells
  See below
• CyTOF-ready
  Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
• Immunoaffinity Purification
  Immobilized anti-His antibody can be used to affinity purify proteins containing exposed His tags. The column capacity must be determined for each individual His-tagged protein.

Western Blot click image to view larger	Detection of His Tag by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line either non-transfected or transfected with His-tagged human CIQ4 and human MYCOC and CHO Chinese hamster ovary cell line transfected with His-tagged human BAI-1. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-His Tag Monoclonal Antibody (Catalog # MAB050) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for His Tag at approximately 32, 65, and 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Flow Cytometry click image to view larger	Detection of His Tag in HEK293 Human Cell Line Transfected with His-tagged Proteins by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with His-tagged proteins was stained with Mouse Anti-His Tag Monoclonal Antibody (Catalog # MAB050, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).
Simple Western click image to view larger	Detection of His Tag by Simple WesternTM. Simple Western lane view shows lysates of CHO Chinese hamster ovary cell line transfected with His-tagged human S1P4/EDG-6, loaded at 0.5 mg/mL. A specific band was detected for His-tagged S1P4/EDG-6 at approximately 75 kDa (as indicated) using 10 µg/mL of Mouse Anti-His Tag Monoclonal Antibody (Catalog # MAB050). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reconstitution
  Reconstitute at 0.5 mg/mL in sterile PBS.

  Reconstitution Buffer Available
• Shipping
  The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
• Stability & Storage
  Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

• Long Name:
  Histidine Tag

• Alternate Names:
  6 His epitope tag; 6X His; 6X-His; H; HHHHHH epitope tag; HHHHHH tag; His Tag; HIS; polyHistidine; Poly-histidine

FAQs

1. This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

   • The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.