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Invitrogen
6x-His Tag Monoclonal Antibody (3D5), AP
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6x-His Tag Antibody (R932-25) in WB
Western blot analysis of Histidine tag (6X His) was performed by loading 20 µg of whole cell extracts of untransfected HEK-293 and HEK-293 transiently overexpressing His- IGFBP6 using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. His-IGFBP6 was detected at ~31 kDa... View More
Product Details
R932-25
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Tag
Published species
Tag
Host/Isotype
Mouse
/ IgG2b
Class
Monoclonal
Type
Antibody
Clone
3D5
Immunogen
6x His synthetic peptide: C-term -His-His-His-His-His-His-COOH
Conjugate
Form
Liquid
Concentration
1-2 mg/mL
Purification
purified
Storage buffer
PBS
Contains
0.01% sodium azide
Storage conditions
Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID
AB_2556555
Product Specific Information
NOTE: R932-25 was previously sold under product #46-0284. When re-ordering, please use #R932-25.
R932-25 detects 6x His tag (C-term) -His-His-His-His-His-His-COOH. Detects fusion proteins containing this (requires the free carboxyl group for detection (Lindner et al., 1997). This product contains enough material for 25 western blots. Western blot experiments with 100 ng of recombinant Positope™ control protein gave a detectable signal within 15 minutes using a chromogenic substrate. The Positope™ control protein is a 53 kDa recombinant protein that contains seven epitope tags, including His(C-term), HisG, c-myc, and V5. For Western blot, do not dilute the AP-conjugated antibody into PBS. Phosphate inhibits alkaline phosphatase. We recommend diluting the AP-conjugated antibody in Tris-Buffered Saline (TBS) containing 0.1% (v/v) Tween-20 and 1% (w/v) nonfat dry milk. If you use a different buffer for washing and blocking your blots, then dilute as described above with that buffer. You may use other blocking agents such as bovine serum albumin (BSA) or gelatin.
All AP-conjugated antibodies have been tested in immunoblotting procedures. Low background was observed. No cross-reactivity has been observed in bacterial lysates.
Target Information
The 6x His tag is a synthetic oligo peptide consisting of 6 consecutive histidine residues (HHHHHH). The His tag is commonly expressed as a tag at either N-or C-terminal regions of recombinant proteins to allow isolation or purification by immobilized metal affinity chromatography. Epitope tagging is a technique in which a known epitope is fused to a recombinant protein using genetic engineering. By choosing a particular epitope and recombinant protein combination, epitope tagging makes it possible to detect proteins for which no antibody is available. His epitope tagged proteins that contain a stretch of histidine residues at the carboxyl or amino terminus are used for purification of recombinant proteins by means of metal chelate chromatography due to metal binding properties of histidine clusters on His tagged proteins. For example, the affinity of the histidine-tag motif to Ni2+ by chelation is strong and selective enough to enable purification of the protein to homogeneity by affinity chromatography on a Ni2+-NTA adsorbent. His tag-specific antibodies are used to facilitate detection or coimmunopreciptation of tagged proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Bioinformatics
Protein Aliases: 6His tag; 6X His; His Tag; Histidine Tag
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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