Anticorps Monoclonal anti-6*His, His-Tag

6*His, His-Tag Monoclonal Antibody for IF/ICC, FC (Intra)

Hôte / Isotype

Mouse / IgG1

Réactivité testée

Protéine recombinante

Applications

IF/ICC, FC (Intra)

Conjugaison

CoraLite® Plus 647 Fluorescent Dye

Publications(7)

CloneNo.

1B7G5

Avis (1)

N° de cat : CL647-66005

Synonymes

6*His, His Tag, His Tag, 6*His, HIS, HHHHHH

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Galerie de données de validation

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  IF Staining of Transfected HEK-293 using CL647-66005

  Immunofluorescent analysis of (-20°C Ethanol) fixed Transfected HEK-293 cells using CoraLite® Plus 647-conjugated 6*His, His-Tag antibody (CL647-66005, Clone: 1B7G5 ) at dilution of 1:500.

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  FC experiment of Transfected HEK-293 using CL647-66005

  1X10^6 Transfected HEK-293 and HEK-293 cells were intracellularly stained with 0.8 ug CoraLite® Plus 647 Anti-N/A 6*His, His-Tag (CL647-66005, Clone:1B7G5).Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Applications testées

• Résultats positifs en IF/ICC: cellules HEK-293 transfectées,
• Résultats positifs en FC (Intra): cellules HEK-293 transfectées,

Dilution recommandée

• Immunofluorescence (IF)/ICC: IF/ICC : 1:250-1:1000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.80 ug per 10^6 cells in a 100 µl suspension

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

• IF: See 3 publications below

Informations sur le produit

CL647-66005 cible 6*His, His-Tag dans les applications de IF/ICC, FC (Intra) 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: 0.84 kDa
• Conjugaison: CoraLite® Plus 647 Fluorescent Dye
• Excitation/Emission maxima wavelengths: 654 nm / 674 nm
• Forme: Liquide
• Méthode de purification: Purification par protéine G
• Tampon de stockage: PBS with 50% glycerol, 0.05% Proclin300, 0.5% BSA
• 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
		Int J Mol Sci; Identifying Exifone as a Dual-Target Agent Targeting Both SARS-CoV-2 3CL Protease and the ACE2/S-RBD Interaction Among Clinical Polyphenolic Compounds; Authors - Jiani Lu
		J Virol; Herpes simplex virus 1 envelope glycoprotein C shields glycoprotein D to protect virions from entry-blocking antibodies; Authors - McKenna A Hull
		bioRxiv; Herpes Simplex Virus 1 Envelope Glycoprotein C Shields Glycoprotein D to Protect Virions from Entry-Blocking Antibodies; Authors - McKenna A Hull
		Synth Syst Biotechnol; Engineered probiotic-mediated intratumoral delivery and controlled release of bacterial collagenase for cancer therapy; Authors - Hong-Rui Li
	IF	ACS Synth Biol; Tumor-Targeted Delivery of PD-1-Displaying Bacteriophages by Escherichia coli for Adjuvant Treatment of Colorectal Cancer; Authors - Hong-Rui Li
	IF	Cell Mol Life Sci; Eeukaryotic-like Sppsk1 from Vibrio splendidus AJ01 mediates phagosome escape via inhibiting phagosome acidification and maturation; Authors - Fa Dai

Avis

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

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FH

Zhi (Verified Customer) (03-22-2024)

Good and easy to use.

• Applications: Immunofluorescence, Flow Cytometry

CL Plus 647 6*His, His-Tag Antibody Immunofluorescence,Flow Cytometry validation ( dilution) in (Cat no:CL647-66005)

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