HIS Tag antibody

Antigen: HIS Tag

Clonality: Monoclonal (HIS-H8)

Host: Mouse

Reactivity: Please enquire

Conjugate: Un-conjugated

Application: Dot Blot (Dot), ELISA, Immunoprecipitation (IP), Immunostaining (ISt), Western Blotting (WB)

Catalog no.: ABIN559683

Additional Information

Immunogen: HHHHHH (6x His) synthetic peptide conjugated to KLH

Isotype: IgG2b

Clone: HIS-H8

Specificity: Recognizes the epitope of 6x His-tags encoded by many commercially available vectors, regardless of the tag's location in the fusion protein sequences (i.e.: reacts with N-terminal, C-terminal or internal 6x His-tags)

Sensitivity: WB (with ECL): 1-2ng of His-tagged fusion proteins

Application Details

Application Notes: WB (with ECL): 1:1000-3000 dilution (incubate for one hour at room temperature) / IS: 1:500-2000 dilution / For best results with other assays (e.g.: Dot, ELISA, IP, etc), please determine optimal working dilution by titration test

Concentration: 1mg/mL

Purification: Protein A affinity chromatography from mouse ascites fluid

Buffer: 10mM PBS, pH 7.2, 0.05% NaN3

Storage: Store at -20°C for up to 2 years or more. Centrifuge after first thaw to maximize product recovery. Avoid repeated freeze-thaw cycles by aliquots. Store aliquots at 4°C for several days to weeks

Restrictions: For Research Use only

Images

LEFT: untransfected control, RIGHT: anti-His (in red) on His-tagged fusion proteins in HEK293 cells. Both counterstained with DAPI (in blue)

Comparison between anti-His tag (HIS.H8 / EH158) mAb with 2 different vendor Abs, probed against a standard ladder (Qiagen Cat. No. 34705) containing five different His-tagged proteins. All Ab dilutions are 1:2000 (0.5μg/mL)

Standard ladder containing five different His-tagged proteins, untransfected control (1), HEK293 cells transfected with His-tagged protein vector (2)

Publications

Product: Zou, Henzel, Liu et al.: "Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3." in: Cell, Vol. 90, Issue 3, pp. 405-13, 1997 (PubMed).

Li, Nijhawan, Budihardjo et al.: "Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade." in: Cell, Vol. 91, Issue 4, pp. 479-89, 1997 (PubMed).

Srinivasula, Ahmad, Fernandes-Alnemri et al.: "Autoactivation of procaspase-9 by Apaf-1-mediated oligomerization." in: Molecular cell, Vol. 1, Issue 7, pp. 949-57, 1998 (PubMed).

Green, Reed: "Mitochondria and apoptosis." in: Science (New York, N.Y.), Vol. 281, Issue 5381, pp. 1309-12, 1998 (PubMed).

Cecconi, Alvarez-Bolado, Meyer et al.: "Apaf1 (CED-4 homolog) regulates programmed cell death in mammalian development." in: Cell, Vol. 94, Issue 6, pp. 727-37, 1998 (PubMed).

Hu, Benedict, Ding et al.: "Role of cytochrome c and dATP/ATP hydrolysis in Apaf-1-mediated caspase-9 activation and apoptosis." in: The EMBO journal, Vol. 18, Issue 13, pp. 3586-95, 1999 (PubMed).

Chi, Bo, Wu et al.: "Vimentin and PSF act in concert to regulate IbeA+ E. coli K1 induced activation and nuclear translocation of NF-κB in human brain endothelial cells." in: PLoS ONE, Vol. 7, Issue 4, pp. e35862, 2012 (PubMed).