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Invitrogen
6x-His Tag (C-term) Monoclonal Antibody (3D5), HRP
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6x-His Tag (C-term) Antibody (R931-25) 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 us... View More
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.jpg?time=20220826?time=20220826 "6x-His Tag (C-term) Antibody (R931-25) in WB")
Product Details
R931-25
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Tag
Published species
C. elegans,
Tag
Host/Isotype
Mouse
/ IgG2b
Class
Monoclonal
Type
Antibody
Clone
3D5
Immunogen
C-term 6x His synthetic peptide: -His-His-His-His-His-His-COOH
Conjugate
Form
Liquid
Purification
purified
Storage buffer
PBS
Contains
no preservative
Storage conditions
Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID
Product Specific Information
NOTE: R931-25 was previously sold under product #46-0707. When re-ordering, please use #R931-25.
This antibody can be used to detect expression of C-terminal His fusion proteins from bacterial, insect, and mammalian cells.
This product contains enough material for 25 Western blots. It is functionally tested using 100 ng of an E. coli expressed fusion protein containing a C-terminal polyhistidine (6xHis) epitope. Chemiluminescent detection should be performed using a 1 minute exposure. For Western blot, dilute in PBS containing 0.05% Tween-20 and 5% nonfat, dry milk (PBSTM).
Using chemiluminescence as the detection method, no cross-reactivity has been observed in bacterial lysates. In mammalian lysates, a few cross-reactive proteins have been observed upon overexposure of blots.
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.
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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