6*His, His-Tag Monoklonaler Antikörper

6*His, His-Tag Monoklonal Antikörper für IF

Wirt / Isotyp

Maus / IgG1

Getestete Reaktivität

rekombinanten Protein und mehr (1)

Anwendung

IF, FC

Konjugation

CoraLite® Plus 488 Fluorescent Dye

Publikationen

3

CloneNo.

1B7G5

Kat-Nr. : CL488-66005

Synonyme

---

Galerie der Validierungsdaten

• 

  IF Staining of Transfected HEK-293 using CL488-66005

  Immunofluorescent analysis of (-20°C Ethanol) fixed Transfected HEK-293 cells using the CoraLite® Plus 488-conjugated version of this antibody, CL488-66005 (6*His, His-Tag antibody), at dilution of 1:100.

---

Geprüfte Anwendungen

• Erfolgreiche Detektion in IF: Transfizierte HEK-293-Zellen

Empfohlene Verdünnung

• Immunfluoreszenz (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

Veröffentlichte Anwendungen

• FC: See 1 publications below

Produktinformation

CL488-66005 bindet in IF, FC 6*His, His-Tag und zeigt Reaktivität mit rekombinanten Protein

• Getestete Reaktivität: rekombinanten Protein
• In Publikationen genannte Reaktivität: human
• Wirt / Isotyp: Maus / IgG1
• Klonalität: Monoklonal
• Typ: Antikörper
• Immunogen: Peptid
• Vollständiger Name: 6*His, His-Tag
• Berechnetes Molekulargewicht: 1 kDa
• Konjugation: CoraLite® Plus 488 Fluorescent Dye
• Excitation/Emission maxima wavelengths: 493 nm / 522 nm
• Form: Liquid
• Reinigungsmethode: Protein-G-Reinigung
• Lagerungspuffer: BS mit 50% Glyzerin, 0,05% Proclin300, 0,5% BSA, pH 7,3.
• 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
		Fish Shellfish Immunol; Apostichopus japonicus matrix metalloproteinase-16 might act as a pattern recognition receptor.; Authors - Jingwei Jiang
human	FC	PLoS Pathog; Breakthrough infection elicits hypermutated IGHV3-53/3-66 public antibodies with broad and potent neutralizing activity against SARS-CoV-2 variants including the emerging EG.5 lineages; Authors - Ling Li
		Fish Shellfish Immunol; A novel short-type peptidoglycan recognition protein with unique polysaccharide recognition specificity in sea cucumber, Apostichopus japonicus; Authors - Jingwei Jiang