AM26687AF-N 6xHistidine Epitope Tag (HHHHHH) antibody

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0.1 mg / €340.00
Please visit the country specific website of Acris Antibodies or contact your local Distributor to buy this product.

Quick Overview

Mouse anti 6xHistidine Epitope Tag 6C4

Product Description for 6xHistidine Epitope Tag

Mouse anti 6xHistidine Epitope Tag 6C4.
Properties: (HHHHHH)
Presentation: Azide Free
Product is tested for Western blot / Immunoblot.

Properties for 6xHistidine Epitope Tag

Product Category Primary Antibodies
Quantity 0.1 mg
Synonyms 6xHis-Tag, HHHHHH Tag, HIS6 Tag, His Tag
Presentation Azide Free
Applications WB
Clonality Monoclonal
Clone 6C4
Host Mouse
Isotype IgG1
Shipping to Not USA/Canada
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Synthetic peptide 6 His
Isotype control AM33027AF-N
Add. information This product was originally produced by MBL International.

Western blot: 1 μg/ml for chemiluminescence detection system. Details see protocol below.
Not recommended for Immunoprecipitation.

Background A variety of plasmids contain DNA that encodes an N-terminal tag consisting of six histidine (His) residues, followed by an extended multiple cloning sites. The His-Tag fusion protein expression system is commonly used because the 6 x His-Tag on the recombinant proteins allows for efficient coupling to Ni++ affinity resins and purification by a single step chromatography.

SDS-PAGE & Western Blotting
1) Mix the sample with equal volume of Laemmli’s sample buffer.
2) Boil the samples for 2 minutes and centrifuge. Load 10 µL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis.
3) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hour in a semi-dry transfer system (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for precise transfer procedure.
4) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4oC.
5) Incubate the membrane with primary antibody diluted with PBS, pH 7.2 containing 1% skimmed milk as suggest in the APPLICATIONS for 1 hour at room temperature. (The concentration of antibody will depend on condition.)
6) Wash the membrane with PBS-T [0.05% Tween-20 in PBS] (5 minutes x 3 times).
7) Incubate the membrane with the 1:10,000 HRP-conjugated anti-mouse IgG diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature.
8) Wash the membrane with PBS-T (5 minutes x 6 times).
9) Wipe excess buffer on the membrane, then incubate it with appropriate chemiluminescence reagent for 1 minute. Remove extra reagent from the membrane by dabbing with paper towel, and seal it in plastic wrap.
10) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. The condition for exposure and development may vary.

General Readings
  1. Porath, J., et al., Protein Express. Purif. 3, 263-281 (1992).

Store (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Shelf life: one year from despatch.

Protein A agarose
Buffer System:
PBS containing 50% glycerol, pH 7.2, without preservatives
Liquid Ig fraction
Azide Free
This specific antibody for His-Tag fusion protein is useful for monitoring of the fusion protein expression and affinity purification.

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