Tested Applications
| Positive IF/ICC detected in | Transfected HEK-293 cells |
| Positive FC (Intra) detected in | Transfected HEK-293 cells |
Recommended dilution
| Application | Dilution |
|---|---|
| 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. | |
Product Information
CL647-66005 targets 6*His, His-Tag in IF/ICC, FC (Intra) applications and shows reactivity with recombinant protein samples.
| Tested Reactivity | recombinant protein |
| Cited Reactivity | human |
| Host / Isotype | Mouse / IgG1 |
| Class | Monoclonal |
| Type | Antibody |
| Immunogen |
Peptide Predict reactive species |
| Full Name | 6*His, His-Tag |
| Calculated Molecular Weight | 0.84 kDa |
| Gene Symbol | |
| Gene ID (NCBI) | |
| RRID | AB_2920271 |
| Conjugate | CoraLite® Plus 647 Fluorescent Dye |
| Excitation/Emission Maxima Wavelengths | 654 nm / 674 nm |
| Excitation Laser | Red Laser (633 nm) |
| Form | Liquid |
| Purification Method | Protein G purification |
| UNIPROT ID | HISTAG |
| 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 -20oC 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.
Protocols
| Product Specific Protocols | |
|---|---|
| FC protocol for CL Plus 647 6*His, His-Tag antibody CL647-66005 | Download protocol |
| IF protocol for CL Plus 647 6*His, His-Tag antibody CL647-66005 | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |
Publications
What published studies show
The 6*His, His-Tag Monoclonal antibody (1B7G5), catalog number CL647-66005, has 11 citations published in journals including Nature Communications, Journal of Extracellular Vesicles, Cell Reports, Cell & Molecular Life Sciences, PLoS Pathogens, ACS Synthetic Biology, Journal of Virology, International Journal of Molecular Sciences, Nature Products Bioprospecting, Synthetic and Systems Biotechnology, and bioRxiv. Research fields span virology (HSV-1, Nipah/Hendra, SARS-CoV-2), bacterial pathogenesis, extracellular vesicle biology, synthetic biology, and cancer therapy.
| Species | Application | Title |
|---|---|---|
J Extracell Vesicles Targeted Intracellular Delivery via Precision Programming of ARRDC1-Mediated Microvesicles. | ||
Nat Commun Highly potent C-type nanoantibodies neutralize Nipah and Hendra viruses by cavity filling on fusion glycoprotein. | ||
Cell Rep Herpes simplex virus type 1 disseminates from the cornea to the CNS in mice by thwarting type III interferon immune defenses. | ||
Cell Mol Life Sci Eeukaryotic-like Sppsk1 from Vibrio splendidus AJ01 mediates phagosome escape via inhibiting phagosome acidification and maturation | ||
Nat Prod Bioprospect Exploring anti-SARS-CoV-2 natural products: dual-viral target inhibition by delphinidin and the anti-coronaviral efficacy of deapio platycodin D | ||
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 |
Reviews
What customers say
The product has received one review with a 5-star rating. The reviewer, Zhi Qiao, used it for Immunofluorescence and Flow Cytometry (lot 21013776) and described it as "good and easy to use," posted on March 22, 2024. Overall, the feedback is positive, highlighting the product's reliability and straightforward usability across multiple applications.
The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.
Zhi (Verified Customer) (03-22-2024) | Good and easy to use.
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