Monoclonal Anti-polyHistidine antibody produced in mouse
SIGMA/SAB4200620 - clone HIS-1, purified immunoglobulin
Synonym: Monoclonal 6 His epitope tag; Monoclonal 6xHis-tag; Monoclonal HHHHHH epitope tag; Monoclonal Hexa His tag; Monoclonal His-tag; Monoclonal His6 tag; Monoclonal Histidine tagged; Monoclonal Poly-His-tag
Product Type: Chemical
Immunofluorescence HEK-293T cells over-expressing N-terminal polyHistidine tagged fusion protein were fixed and permeabilized with 4% paraformaldehyde followed by 0.5% Triton X-100. Fixed cells were stained with 8 μg/mL Monoclonal Anti-polyHistidine, Clone: HIS-1 (Cat. No. SAB4200620). The antibody was developed using Goat Anti-Mouse IgG, Cy3™ conjugate. Cells were counterstained with DAPI (blue) to stain nuclei.
Immunoblotting Recombinant multi-tag protein produced in E. coli with C-terminal polyHistidine was separated on SDS-PAGE and probed with Monoclonal Anti-polyHistidine, Clone: HIS-1 (Cat. No. SAB4200620). The antibody was developed using Goat Anti-Mouse IgG-Peroxidase (Cat. No. A2304) and a chemiluminescent substrate. Lanes 1. 0.5 μg/mL antibody 2. 0.25 μg/mL antibody 3. 0.125 μg/mL antibody 4. Negative control: without first antibody
Flow Cytometry
1×106 human HEK-293T cells over-expressing C-terminal HIS tagged fusion protein were fixed and permeabilized with 2% Paraformaldehyde followed by 0.1% Triton X-100. Fixed cells were labeled with 6 μg/test Monoclonal Anti-polyHistidine Clone:HIS-1 (Cat. No. SAB4200620) followed by Rabbit Anti-Mouse IgG, FITC conjugate. Green histogram: negative control without first antibody, Purple histogram: antibody specific staining.
1×106 human HEK-293T cells over-expressing C-terminal HIS tagged fusion protein were fixed and permeabilized with 2% Paraformaldehyde followed by 0.1% Triton X-100. Fixed cells were labeled with 6 μg/test Monoclonal Anti-polyHistidine Clone:HIS-1 (Cat. No. SAB4200620) followed by Rabbit Anti-Mouse IgG, FITC conjugate. Green histogram: negative control without first antibody, Purple histogram: antibody specific staining.
| antibody form | purified immunoglobulin |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | HIS-1, monoclonal |
| concentration | ~1.0 mg/mL |
| isotype | IgG2a |
| packaging | antibody small pack of 25 μL |
| Quality Level | 200  |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | flow cytometry: 3-6 μg/test |
| immunoblotting: 0.25-0.5 μg/mL | |
| immunofluorescence: 4-8 μg/mL |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | Monoclonal Anti-polyHistidine (mouse IgG2a isotype) is derived from the HIS-1 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from a BALB/c mouse immunized with a recombinant His-tagged fusion protein. The antibody is purified from culture supernatant of hybridoma cells grown in a bioreactor. Monoclonal anti-polyHistidine recognizes native as well as denatured, reduced forms of synthetic polyHistidine or polyhistidine-tagged fusion proteins. The product is reactive with fusion protein expressed by prokaryotic pET, pRSET, and pTrc expression vectors. The antibody preferentially recognizes N-terminal tagged fusion proteins. |
| Other Notes: | In order to obtain the best results using various techniques and preparations, we recommend determining optimal working dilutions by titration. Use of sensitive film is recommended. |
| Physical form: | Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide as a preservative. |
| Preparation Note: | For extended storage, freeze at -20 °C in working aliquots. Repeated freezing and thawing or storage in “frost-free” freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |