6*His, His-Tag Monoclonal antibody

6*His, His-Tag Monoclonal Antibody for IF, IP, WB, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

recombinant protein

Applications

WB, IP, IF, FC, CoIP, ELISA

Conjugate

Unconjugated

CloneNo.

1B7G5

Cat no : 66005-1-Ig

Synonyms

6*His, 6*His, His Tag, 6histag, 6XHis, HHHHHH, HIS, His probe, His Tag, His-probe, HISTIDINE TAGGED



Tested Applications

Positive WB detected inrecombinant protein
Positive IP detected inTransfected HEK-293 cells
Positive IF detected inTransfected HEK-293 cells
Planning an IF experiment? We recommend our CoraLite®488 and CoraLite®594 conjugated versions of this antibody.

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:5000-1:50000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for IP and 1:5000-1:50000 for WB
Immunofluorescence (IF)IF : 1:200-1:800
Sample-dependent, check data in validation data gallery

Product Information

66005-1-Ig targets 6*His, His-Tag in WB, IP, IF, FC, CoIP, ELISA applications and shows reactivity with recombinant protein samples.

Tested Reactivity recombinant protein
Host / Isotype Mouse / IgG1
Class Monoclonal
Type Antibody
Immunogen Peptide
Full Name 6*His, His-Tag
Calculated molecular weight 1 kDa
Gene symbol
Gene ID (NCBI)
Conjugate Unconjugated
Form Liquid
Purification Method Protein G purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.

Background Information

Protein tags are a protein or peptide sequences located either on the C- or N- terminal of the target protein. His-tag is often used for affinity purification and binding assays. Expressed. The His-tag antibody is a useful tool for monitoring of the His-tagged proteins and recognizes His-tags placed at N-terminal, C-terminal, and internal regions of fusion proteins expressed in bacteria, insect, and mammalian cells. A His-tag (polyhistidine tag) consists of at least six histidine residues that are located at the N- or C-terminus of recombinant proteins. It is commonly used for affinity purification and protein binding experiments.

 

The recombinant protein has only 6 histidine residues as a tag. Is this sufficient to be detected by your antibody? Is the His-tag antibody able to detect N-terminal and C-terminal His tags?

The His-tag antibody was raised using a 6-His tag peptide as the immunogen, which is ~1 kDa. Some scientists use more than 6 histidine residues as a tag for recombinant proteins to increase affinity to metal ions. A 6-histidine tag is sufficient for specific recognition by the His-tag antibody. It is able to recognize both N-terminal and C-terminal His tags of recombinant proteins produced in various expression systems, including bacteria, yeasts, insect, and mammalian cells.

 

After purification, I can detect my protein by western blotting using the His-tag antibody but can also see additional bands running lower. What might they represent?

They may represent cleavage products of the recombinant protein. Unspecific protein cleavage can occur during protein production (e.g., in bacteria) or protein purification post the cell lysis steps. Changing expression conditions (temperature, time, induction methods, medium) and using protease inhibitors during purification helps to minimize protein degradation. Additional steps, such as ion exchange, size exclusion, or chromatography, allow isolation of the full-length product.

 

Can a His-tag antibody be used in applications other than western blotting?

Yes, this antibody has been successfully used for immunofluorescence (IF), immunoprecipitation (IP), and indirect ELISA analysis.


Protocols

Product Specific Protocols
IF protocol for 6*His, His-Tag antibody 66005-1-IgDownload protocol
IP protocol for 6*His, His-Tag antibody 66005-1-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
WB

Oncol Rep

SOCS4 expressed by recombinant HSV protects against cytokine storm in a mouse model.

Authors - Shuqi Ren

Hum Mol Genet

GRP75 overexpression rescues frataxin deficiency and mitochondrial phenotypes in Friedreich Ataxia cellular models.

Authors - Yi Na Dong

Elife

TRIM28 promotes HIV-1 latency by SUMOylating CDK9 and inhibiting P-TEFb.

Authors - Xiancai Ma

Fish Shellfish Immunol

Vibrio parahaemolyticus flagellin induces cytokines expression via toll-like receptor 5 pathway in orange-spotted grouper, Epinephelus coioides.

Authors - Liangge He

Cell Rep

The EF-Hand Protein CALML6 Suppresses Antiviral Innate Immunity by Impairing IRF3 Dimerization.

Authors - Ziyang Wang

J Virol

GADD45γ activated early in the course of HSV-1 infection suppresses the activation of a network of innate immunity genes.

Authors - Mingmin She

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.


FH

Adrian (Verified Customer) (04-22-2020)

We tested anti-His tag Ab's in our immunometric analyses using recombinant his-tagged nickel binding protein NikR as a primary Ab. We had no issue obtaining a signal, but the paper eventually was left out of this part because our primary Ab had stability issues. It was a few years ago, though.

  • Applications: Western Blot
  • Cell Tissue Type: in vitro studies on sybthetic nickel-chelate complex binding
FH

Paul (Verified Customer) (01-15-2020)

Works well for Western Blot.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: HEK293T
FH

Laura (Verified Customer) (01-14-2020)

Good sensitivity for Western blot.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: HEK293T
FH

Aamir (Verified Customer) (01-08-2020)

Works well for WB

  • Applications: Western Blot,
  • Primary Antibody Dilution: 5000
  • Cell Tissue Type: HEKs
FH

Benjamin (Verified Customer) (01-07-2020)

Works very well. Detects a his-tagged fusion protein with high sensitivity and very little background in western blot.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: HEK lysate and recombinant his tagged protein
FH

Nikhil (Verified Customer) (10-16-2019)

Recombinant His tagged p53 expressed in Bacteria. Lane 1 marker, Lane2-lane 6: Elutions 1-5

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
6*His, His-Tag Antibody Western Blot, validation (1:1000 dilution) in (Cat no:66005-1-Ig)
FH

Shashirekha (Verified Customer) (07-09-2019)

The antibody is good but it stains ladder as well.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: Mouse spinal cord
6*His, His-Tag Antibody Western Blot, validation (1:5000 dilution) in Mouse spinal cord (Cat no:66005-1-Ig)
FH

Chun (Verified Customer) (07-03-2019)

This antibody is excellent.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:2,000
  • Cell Tissue Type: HEK293 and HeLa cells
FH

John (Verified Customer) (02-27-2019)

We unfortunately saw unexpected fluorescence in negative controls when using this antibody, so were unable to draw conclusions from our initial experiments. On a more positive note, after running through attempts to resolve the issue Proteintech did offer us an alternative antibody to try.

  • Applications: Immunofluorescence,
  • Primary Antibody Dilution: 1:40 and 1:200
  • Cell Tissue Type: SH-SY5Y
FH

Marisa (Verified Customer) (02-05-2019)

Highly dependent on sample type. Works very nice and shurly up to 1:50,000 detecting recombinant His-proteins of E. coli. Difficult to analyze proteins produced in insect cells. Please use 1:1000 as a dilution.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:5000
FH

Youjun (Verified Customer) (11-27-2018)

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: HEK293 cells