6*His, His-Tag Monoklonaler Antikörper

6*His, His-Tag Monoklonal Antikörper für IF/ICC, FC (Intra)

Wirt / Isotyp

Maus / IgG1

Getestete Reaktivität

rekombinanten Protein

Anwendung

IF/ICC, FC (Intra)

Konjugation

CoraLite® Plus 647 Fluorescent Dye

Publikationen(7)

CloneNo.

1B7G5

Rezensionen (1)

Kat-Nr. : CL647-66005

Synonyme

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

---

• 

  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.

• 

  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).

---

Geprüfte Anwendungen

• Erfolgreiche Detektion in IF/ICC: Transfizierte HEK-293-Zellen
• Erfolgreiche Detektion in FC (Intra): Transfizierte HEK-293-Zellen

Empfohlene Verdünnung

• Immunfluoreszenz (IF)/ICC: IF/ICC : 1:250-1:1000
• Durchflusszytometrie (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

Veröffentlichte Anwendungen

• IF: See 3 publications below

Produktinformation

CL647-66005 bindet in IF/ICC, FC (Intra) 6*His, His-Tag und zeigt Reaktivität mit rekombinanten Protein

• Getestete Reaktivität: rekombinanten Protein
• Wirt / Isotyp: Maus / IgG1
• Klonalität: Monoklonal
• Typ: Antikörper
• Immunogen: Peptid
• Vollständiger Name: 6*His, His-Tag
• Berechnetes Molekulargewicht: 0.84 kDa
• Konjugation: CoraLite® Plus 647 Fluorescent Dye
• Excitation/Emission maxima wavelengths: 654 nm / 674 nm
• Form: Liquid
• Reinigungsmethode: Protein-G-Reinigung
• Lagerungspuffer: PBS with 50% glycerol, 0.05% Proclin300, 0.5% BSA
• Lagerungsbedingungen: Bei -20°C lagern. Vor Licht schützen. Nach dem Versand ein Jahr stabil. Aliquotieren ist bei -20^oC Lagerung nicht notwendig. 20ul Größen enthalten 0,1% BSA.

Hintergrundinformationen

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.

Publikationen

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

Rezensionen

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

---

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)