Anticorps Monoclonal anti-6*His, His-Tag

6*His, His-Tag Monoclonal Antibody for IF

Hôte / Isotype

Mouse / IgG1

Réactivité testée

Protéine recombinante

Applications

WB, IF

Conjugaison

CoraLite®594 Fluorescent Dye

Publications

1

CloneNo.

1B7G5

N° de cat : CL594-66005

Synonymes

6*his tag

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Galerie de données de validation

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  IF Staining of Transfected HEK-293 using CL594-66005

  Immunofluorescent analysis of (-20°C Ethanol) fixed Transfected HEK-293 cells using the CoraLite®594-conjugated version of this antibody, CL594-66005 (6*His, His-Tag antibody), at dilution of 1:100.

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Applications testées

• Résultats positifs en IF: cellules HEK-293 transfectées,

Dilution recommandée

• Immunofluorescence (IF): IF : 1:50-1:500

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

Applications publiées

• WB: See 1 publications below

Informations sur le produit

CL594-66005 cible 6*His, His-Tag dans les applications de WB, IF et montre une réactivité avec des échantillons Protéine recombinante

• Réactivité: Protéine recombinante
• Hôte / Isotype: Mouse / IgG1
• Clonalité: Monoclonal
• Type: Anticorps
• Immunogène: Peptide
• Nom complet: 6*His, His-Tag
• Masse moléculaire calculée: 1 kDa
• Conjugaison: CoraLite®594 Fluorescent Dye
• Excitation/Emission maxima wavelengths: 588 nm / 604 nm
• Forme: Liquide
• Méthode de purification: Purification par protéine G
• Tampon de stockage: PBS avec glycérol à 50 %, Proclin300 à 0,05 % et BSA à 0,5 %, pH 7,3.
• Conditions de stockage: Stocker à -20 °C. Éviter toute exposition à la lumière. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

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.

Publications

Species	Application	Title
	WB	Cell Rep; Structural basis for translation inhibition by MERS-CoV Nsp1 reveals a conserved mechanism for betacoronaviruses; Authors - Swapnil C Devarkar