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6X His-tag, AlpHcAbs Rabbit Alpaca Monoclonal Antibody

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Catalog# HA721172

6X His-tag, AlpHcAbs Rabbit Alpaca Monoclonal Antibody

Application

  • WB

  • IP

  • ELISA

Reactivity
Conjugation

  • unconjugated

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

Overview

Product Name

6X His-tag, AlpHcAbs Rabbit Alpaca Monoclonal Antibody

Antibody Type

Alpaca Nanobody

Immunogen

6*His tag fusion protein.

Validated Applications

WB, IP, ELISA

Positive Control

C-terminal His-tag fusion protein lysate, N-terminal His-tag fusion protein lysate, Mid His-tag fusion protein lysate.

Conjugation

unconjugated

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

PBS pH=7.5

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:10,000-1:50,000 (C-His)

  • WB

  • 1:500-1:10,000 (N-His)

  • WB

  • 1:10,000-1:50,000 (Mid-His)

  • IP

  • 1:100-1:500

  • ELISA

  • 1:10,000-1:50,000

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. Anti-His tag, AlpHcAbs Rabbit antibody is monovalentrecom mbinant single domain antibody fused to rabbit 1gG Fc,and Anti-His tag AlpHcAbs Rabbit antibody detects the His tag selectively,no re eactivity with other proteins.

Background References

1. Hengen, Paul N (1995). Purification of His-Tag fusion proteins from Escherichia coli. Trends in Biochemical Sciences. 20 (7): 285–6.

2. Hochuli, E.; Döbeli, H.; Schacher, A. (January 1987). New metal chelate adsorbent selective for proteins and peptides containing neighbouring histidine residues. Journal of Chromatography A. 411: 177–184.

Synonyms

6 His epitope tag antibody

Hexa His tag antibody

HHHHHH epitope tag antibody

HHHHHH tag antibody

His tag antibody

Polyhistidine Tag antibody

Images

  • Western blot analysis of 6X His-tag, AlpHcAbs Rabbit on different lysates with Rabbit anti-6X His-tag, AlpHcAbs Rabbit antibody (<a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a>) at 1/50,000 dilution.<br /><br />Lane 1: C-terminal His-tag fusion protein lysate<br />Lane 2: N-terminal His-tag fusion protein lysate<br />Lane 3/4/5/6: Mid His-tag fusion protein lysate<br /><br />Lysates/proteins at 50 ng/Lane.<br /><br />Observed band size: 30/45/50 kDa<br /><br />Exposure time: 30 seconds;<br /><br />12% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a>) at 1/50,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of 6X His-tag, AlpHcAbs Rabbit on different lysates with Rabbit anti-6X His-tag, AlpHcAbs Rabbit antibody (HA721172) at 1/50,000 dilution.

    Lane 1: C-terminal His-tag fusion protein lysate
    Lane 2: N-terminal His-tag fusion protein lysate
    Lane 3/4/5/6: Mid His-tag fusion protein lysate

    Lysates/proteins at 50 ng/Lane.

    Observed band size: 30/45/50 kDa

    Exposure time: 30 seconds;

    12% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721172) at 1/50,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • His-tag was immunoprecipitated in 2µg C-terminal His-tag fusion protein lysate with <a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a>. Western blot was performed from the immunoprecipitate using <a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a> at 1/10,000 dilution. Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 60 mins at room temperature.<br /><br />Lane 1: C-terminal His-tag fusion protein lysate (input).<br />Lane 2: <a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a> IP in C-terminal His-tag fusion protein lysate.<br />Lane 3: Rabbit IgG instead of <a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a> IP in C-terminal His-tag fusion protein lysate.<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST.

    His-tag was immunoprecipitated in 2µg C-terminal His-tag fusion protein lysate with HA721172. Western blot was performed from the immunoprecipitate using HA721172 at 1/10,000 dilution. Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 60 mins at room temperature.

    Lane 1: C-terminal His-tag fusion protein lysate (input).
    Lane 2: HA721172 IP in C-terminal His-tag fusion protein lysate.
    Lane 3: Rabbit IgG instead of HA721172 IP in C-terminal His-tag fusion protein lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • His-tag was immunoprecipitated in 2µg N-terminal His-tag fusion protein lysate with <a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a>. Western blot was performed from the immunoprecipitate using <a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a> at 1/10,000 dilution. Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 60 mins at room temperature.<br /><br />Lane 1: N-terminal His-tag fusion protein lysate (input).<br />Lane 2: <a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a> IP in N-terminal His-tag fusion protein lysate.<br />Lane 3: Rabbit IgG instead of <a href="/products/HA721172" style="font-weight: bold;text-decoration: underline;">HA721172</a> IP in N-terminal His-tag fusion protein lysate.<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST.

    His-tag was immunoprecipitated in 2µg N-terminal His-tag fusion protein lysate with HA721172. Western blot was performed from the immunoprecipitate using HA721172 at 1/10,000 dilution. Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 60 mins at room temperature.

    Lane 1: N-terminal His-tag fusion protein lysate (input).
    Lane 2: HA721172 IP in N-terminal His-tag fusion protein lysate.
    Lane 3: Rabbit IgG instead of HA721172 IP in N-terminal His-tag fusion protein lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST.

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

Citation