Anticorps Monoclonal anti-6*His, His-Tag

6*His, His-Tag Monoclonal Antibody for WB

Hôte / Isotype

Mouse / IgG1

Réactivité testée

Protéine recombinante

Applications

WB

Conjugaison

CoraLite® Plus 750 Fluorescent Dye

CloneNo.

1B7G5

N° de cat : CL750-66005

Synonymes

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Galerie de données de validation

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  WB analysis using CL750-66005

  Various lysates were subjected to SDS PAGE followed by western blot with CL750-66005 (6*His, His-Tag antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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Applications testées

• Résultats positifs en WB: Transfected HEK-293T cells,

Dilution recommandée

• Western Blot (WB): WB : 1:1000-1:4000

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

Informations sur le produit

CL750-66005 cible 6*His, His-Tag dans les applications de WB 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® Plus 750 Fluorescent Dye
• Excitation/Emission maxima wavelengths: 755 nm / 780 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.