Anti-6-His Epitope Tag Affinity Matrix

Pricing & Availability

Clone: 6-His (See other available formats)
Regulatory Status: RUO
Other Names: 6-His Epitope Tag
Previously: Covance Catalog# AFC-156P
Isotype: Mouse IgG1, κ
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Product Citations: publications

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900501

1 mL

$633

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Description

Monoclonal mouse antibody 6-His recognizes the peptide epitope, HHHHHH. This antibody was purified using protein-G chromatography and was subsequently immobilized onto a Sepharose™ Fast Flow matrix.

Product Details

Technical data sheet

Product Details

Verified Reactivity: Epitope tag
Reported Reactivity: Species independent
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Monoclonal 6-His antibody was raised against a recombinant protein containing the sequence HHHHHH.
Formulation: Purified IgG immobilized on Sepharose™ Fast Flow beads (in PBS + 0.03% Thimerosal).
Preparation: The antibody was purified by affinity chromatography.
Storage & Handling: Store between 2-8°C. The matrix may be re-used several times. To strip column after use, wash with several bead volumes of 0.1 M glycine pH 2.8 followed immediately by PBS containing 0.3% Thimerosal or 1mM Sodium Azide as preservative.
Application: Purification, IP
Recommended Usage: Each lot of this antibody is quality control tested by immunoprecipitation.

The optimal buffers and matrix concentration should be determined for each specific assay condition.

Binding: Tagged protein will bind to matrix in common physiologic buffers with pH in the range of 6.0-7.5, salt from 50-500 mM and in the presence of reasonable levels of detergent. Excess reducing agent should be avoided as the disulfide bridges holding antibody heavy and light chains may be compromised. BioLegend tests Affinity Matrix using an equilibration/binding buffer containing:
• 100 mM Tris-HCl (pH 7.5)
• 150 mM NaCl
• 0.1% Tween 20
• 0.5% BSA
• 1 mM beta-mercaptoethanol

Washing: After binding, washes with several bead volumes of buffer are recommended. Such buffer may contain increased salt, altered pH, etc. as determined empirically to remove un-tagged proteins.

Elution: Several options are available for elution.
1. SDS gel loading buffer may be applied directly to the beads in order to display all bound protein on a polyacrylamide gel/western. Note that gel loading buffer containing reducing agent will also release some antibody heavy and light chains (approx 25 and 50kD, respectively).
2. Competitive elution with epitope peptide. For epitope tag affinity matrices, prepare an elution buffer with epitope tag peptide at 400 ug/mL in 50 mM Tris-HCl (pH 7.5), 50 mM NaCl, 1 mM EDTA (pH 8.0).
3. Chemical Elution. Elution by pH or chaotropic salts is also possible. For elution by pH, either 0.1 M glycine pH 2.8 or 40 mM diethyl-amine pH 11.0 may be used.
Application Notes: This affinity matrix can be used for immunopurification of 6-His-tagged fusion proteins from crude starting material. On an analytical scale, it can also be used for immunoprecipitating His-tagged proteins.

Sepharose is a trademark of GE Life Sciences
RRID: AB_2564996 (BioLegend Cat. No. 900501)

Antigen Details

Biology Area: Cell Biology
Gene ID: NA
UniProt: View information about 6-His on UniProt.org

Related Products

Description	Clone	Applications

Other Formats

View All 6-His Reagents Request Custom Conjugation

Description	Clone	Applications
Anti-6-His Epitope Tag Affinity Matrix	6-His	IP,Purification
Biotin anti-6-His Epitope Tag	6-His	WB,ICC,IP
FITC anti-6-His Epitope Tag	6-His	ICC
Purified anti-6-His Epitope Tag	6-His	WB,ICC,IP
Direct-Blot™ HRP anti-6-His Epitope Tag	6-His	WB
Alexa Fluor® 647 anti-6-His Epitope Tag	6-His	ICC

Certificate of Analysis

Go To Top Version: 7 Revision Date: 09/19/2023

For Research Use Only. Not for diagnostic or therapeutic use.

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

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