Western blot analysis of HisG was performed by loading 20 µg of whole cell extracts of untransfected HEK-293 cell lysate (lane1) and HEK-293 transiently expressing HisG-RPS6 (lane2) 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 atleast 1 hour at room temperature. HisG was detected at ... View More

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Product Details

R942-25

Applications

Tested Dilution

Publications

Western Blot (WB)

1:2,000

Product Specifications

Species Reactivity

Tag

Host/Isotype

Mouse / IgG2a

Class

Monoclonal

Type

Antibody

Immunogen

6x His synthetic peptide: -His-His-His-His-His-His-Gly

Conjugate

AP AP AP

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.

AB_2556559

Product Specific Information

NOTE: R942-25 was previously sold under product #46-0286. When re-ordering, please use #R942-25.

This antibody is designed to specifically detect recombinant proteins containing the seven amino acid 6xHis-Gly epitope. This epitope is present in many expression vectors encoding an N-terminal 6xHis tag. The HisG antibody recognizes the following sequence: -His-His-His-His-His-His-Gly.

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, we recommend diluting the AP-conjugated antibody 1:2000 into Tris-Buffered Saline (TBS) containing 0.1% (v/v) Tween-20 and 1% (w/v) nonfat dry milk.

Target Information

Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Expression of recombinant proteins in E. coli as a fusion protein with neighboring histidine residues is one of the most popular methods of epitope tags. 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. HisG antibodies allow detection of recombinant proteins containing a polyhistidine sequence: His-His-His-His-His-His-Gly (6xHis-Gly epitope). HisG antibodies can be used to detect expression of fusion proteins from bacterial, insect, and mammalian cells.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: 6xHis-Gly; 6xHis-Glycine; His G; His Tag; Histidine Tag

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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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HisG Epitope Tag Monoclonal Antibody, AP

HisG Epitope Tag Antibody (R942-25) in WB