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6X His tag Mouse Monoclonal Antibody [A5C12]

Usd: 350

Specification

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Catalog# HA600028

6X His tag Mouse Monoclonal Antibody [A5C12]

Application

  • WB

  • ELISA

Reactivity

  • Species independent

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

6X His tag Mouse Monoclonal Antibody [A5C12]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide HHHHHHGCGHHHHHH.

Species Reactivity

Species independent

Validated Applications

WB, ELISA

Positive Control

C-terminal His-tagged recombinant protein lysates, N-terminal His-tagged recombinant protein lysates.

Conjugation

unconjugated

Clone Number

A5C12

RRID

AB_3071645

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:10,000

  • ELISA

  • Use at an assay dependent concentration.

Target

Function

His-Tag Antibody is a high quality monoclonal His-Tag antibody (also designated 6X His-Tag antibody or His-Probe antibody) suitable for the detection of the His-Tag protein. His-Tag Antibody is available as both the non-conjugated anti-His-Tag antibody form, as well as multiple conjugated forms of anti-His-Tag antibody, including agarose, HRP, PE, FITC and multiple Alexa Fluor® conjugates. Plasmid vectors for the expression of coding regions of eukaryotic genes in bacterial, insect and mammalian hosts are in common usage; such expression vectors are frequently used to encode hybrid fusion proteins consisting of a eukaryotic target protein and a specialized region designed to aid in the purification and visualization of the target protein. A system that has proven to be very successful relies on the insertion of a six histidine (His6) sequence in the N-terminus of the encoded protein, allowing for efficient coupling to Ni2+-chelating resins and purification by single step affinity chromatography. This polyhistidine sequence can then be removed by specific cleavage at sites recognized by enzymes such as thrombin or enterokinase, permitting the separation of the target protein from the polyhistidine tag. Visualization of such fusion proteins can be achieved by utilizing antibodies generated against specific peptide sequences downstream from the multiple cloning site.

Background References

1. Meng L. et. al. Effects of His-tag on Catalytic Activity and Enantioselectivity of Recombinant Transaminases. Appl Biochem Biotechnol. 2020 Mar

Images

  • Western blot analysis of His-tag on C-terminal His-tagged recombinant protein lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/HA600028" style="font-weight: bold;text-decoration: underline;">HA600028</a>, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:20,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of His-tag on C-terminal His-tagged recombinant protein lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (HA600028, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of His-tag on N-terminal His-tagged recombinant protein lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/HA600028" style="font-weight: bold;text-decoration: underline;">HA600028</a>, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:20,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of His-tag on N-terminal His-tagged recombinant protein lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (HA600028, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.

  • Immobilized His-tagged recombinant protein at 1 μg/ml overnight at 4℃. Then blocked with 1% BSA for 1 hour at 37℃, and incubated with the primary antibody for 1 hour at 25℃.

    Immobilized His-tagged recombinant protein at 1 μg/ml overnight at 4℃. Then blocked with 1% BSA for 1 hour at 37℃, and incubated with the primary antibody for 1 hour at 25℃.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation