CoraLite® Plus 488-conjugated 6*His, His-Tag Monoclonal antibody

6*His, His-Tag Monoclonal Antibody for IF/ICC

Host / Isotype

Mouse / IgG1

Reactivity

recombinant protein

Applications

IF/ICC

Conjugate

CoraLite® Plus 488 Fluorescent Dye

Publications

3

CloneNo.

1B7G5

Cat no : CL488-66005

Synonyms

6*His, His Tag, His probe, HIS, HHHHHH, 6XHis

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Validation Data Gallery

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  IF Staining of Transfected HEK-293 using CL488-66005

  Immunofluorescent analysis of (-20°C Ethanol) fixed Transfected HEK-293 cells using CoraLite® Plus 488 6*His, His-Tag antibody (CL488-66005, Clone: 1B7G5 ) at dilution of 1:200.

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Tested Applications

• Positive IF/ICC detected in: Transfected HEK-293 cells

Recommended dilution

• Immunofluorescence (IF)/ICC: IF/ICC : 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.

Product Information

CL488-66005 targets 6*His, His-Tag in IF/ICC applications and shows reactivity with recombinant protein samples.

• Tested Reactivity: recombinant protein
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: 6*His, His-Tag
• Calculated Molecular Weight: 1 kDa
• RRID: AB_2883223
• Conjugate: CoraLite® Plus 488 Fluorescent Dye
• Excitation/Emission Maxima Wavelengths: 493 nm / 522 nm
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 50% Glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3.
• Storage Conditions: Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

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
		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; Apostichopus japonicus matrix metalloproteinase-16 might act as a pattern recognition receptor.; Authors - Jingwei Jiang
		Fish Shellfish Immunol; A novel short-type peptidoglycan recognition protein with unique polysaccharide recognition specificity in sea cucumber, Apostichopus japonicus; Authors - Jingwei Jiang