6*His, His-Tag Monoclonal antibody

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

Host / Isotype

Mouse / IgG1

Reactivity

recombinant protein and More (5)

Applications

WB, IP, IHC, IF, FC, CoIP, ChIP, Cell treatment, ELISA

Conjugate

Unconjugated

CloneNo.

1B7G5

Cat no : 66005-1-Ig

Synonyms

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


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  • Western Blot (WB) analysis of recombinant protein using 6*His, His-Tag Monoclonal antibody (66005-1-Ig)

    WB analysis of recombinant protein using 66005-1-Ig

    Western blot of 6*His-tagged fusion protein with anti-6*HIS tag (66005-1-Ig) at various dilutions.

  • Immunoprecipitation (IP) experiment of Transfected HEK-293 cells using 6*His, His-Tag Monoclonal antibody (66005-1-Ig)

    IP experiment of Transfected HEK-293 using 66005-1-Ig

    IP Result of anti-6*His, His-Tag (IP:66005-1-Ig, 7ug; Detection:66005-1-Ig 1:10000) with Transfected HEK-293 cells lysate 300ug.

  • Immunofluorescence (IF) / fluorescent staining of Transfected HEK-293 cells using 6*His, His-Tag Monoclonal antibody (66005-1-Ig)

    IF Staining of Transfected HEK-293 using 66005-1-Ig

    Immunofluorescent analysis of (-20°C Ethanol) fixed Transfected HEK-293 cells using 66005-1-Ig (6*His, His-Tag antibody) at dilution of 1:400 and Alexa Fluor 594-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

"6*His, His-Tag Antibodies" Comparison

View side-by-side comparison of 6*His, His-Tag antibodies from other vendors to find the one that best suits your research needs.

Tested Applications

Positive WB detected inrecombinant protein
Positive IP detected inTransfected HEK-293 cells
Positive IF detected inTransfected HEK-293 cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:5000-1:50000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunofluorescence (IF)IF : 1:200-1:800
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Product Information

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

Tested Reactivity recombinant protein
Cited Reactivityhuman, mouse, chicken, yeast, pig
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)
RRIDAB_11232599
Conjugate Unconjugated
Form Liquid
Purification MethodProtein A 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. 20ul sizes contain 0.1% BSA.

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

Nature

IGF1R is an entry receptor for respiratory syncytial virus.

Authors - Cameron D Griffiths

Nature

UDP-glucose accelerates SNAI1 mRNA decay and impairs lung cancer metastasis.

Authors - Xiongjun Wang
WB

Nature

CRISPR screens unveil signal hubs for nutrient licensing of T cell immunity.

Authors - Lingyun Long
WB

Science

Structural insight into the SAM-mediated assembly of the mitochondrial TOM core complex.

Authors - Qiang Wang
WB

Nature

Fatal swine acute diarrhoea syndrome caused by an HKU2-related coronavirus of bat origin.

Authors - Peng Zhou

Cell

LECT2, a Ligand for Tie1, Plays a Crucial Role in Liver Fibrogenesis.

Authors - Meng Xu

Reviews

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


FH

Raquel (Verified Customer) (04-13-2023)

Nice selective staining. No background detected when looking for our 6xHis tagged recombinant protein within yeast supernatant. Good quality western blot image. Great value for money and would recommend.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:10,000
  • Cell Tissue Type: Yeast supernatant.
FH

Ken (Verified Customer) (01-30-2023)

This is a specific antibody. It detects His-tagged proteins expressed in 293T cells with low background.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:2000
  • Cell Tissue Type: His-tagged protein expressed in human tissue culture cells
FH

Samruddhi (Verified Customer) (02-22-2022)

The antibody works well for Western Blot, and there are no unspecific bands.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:2000
FH

Amie (Verified Customer) (07-27-2021)

I ran two identical gels, with two different sized His-tagged proteins on them, and some lanes that should contain no His-tagged protein. As you can see in the image, the Proteintech antibody had much more background and less specificity than the anti-His antibody I currently use (I cannot recall what vendor it is from). It is not useful for my purposes given the cross-reactivity/lack of specificity.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: his-tagged proteins expressed in yeast
6*His, His-Tag Antibody Western Blot validation (1:5000 dilution) in his-tagged proteins expressed in yeast (Cat no:66005-1-Ig)
FH

Barry (Verified Customer) (01-07-2021)

Suitable

  • Applications: Flow Cytometry
  • Primary Antibody Dilution: n/a
  • Cell Tissue Type: Cell Type
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