Western blot analysis of E. coli lysate containing 6-His-tagged protein with anti-6-His-tag antibody. Western blot analysis was performed on 160 ng, 80 ng or 20 ng of E. coli lysate containing 6-His-tagged protein. Cells were lysed with SDS sample buffer. The PVDF membrane was blocked with 5% dry milk in PBST for 1 h at RT. The primary antibody anti-6-His-tag mouse monoclonal, 6His was diluted in blocking buffer (antibody concentration 0.125 µg/ml) and incubated for 1 h at RT. The secondary antibody goat anti-mouse IgG polyclonal, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Western blot analysis of E. coli lysate containing 6-His-tagged protein with anti-6-His-tag antibody. Western blot analysis was performed on 50 ng or 25 ng of E. coli lysate containing 6-His-tagged protein. Cells were lysed with SDS sample buffer. The PVDF membrane was blocked with 5% dry milk in PBST for 1 h at RT. The primary antibody anti-6-His-tag mouse monoclonal, 6His was diluted in blocking buffer (antibody concentration 0.05 µg/ml) and incubated for 1 h at RT. The secondary antibody goat anti-mouse IgG polyclonal, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Western blot analysis of E. coli lysate containing 6-His-tagged protein with anti-6-His-tag antibody. Western blot analysis was performed on 160 ng, 80 ng or 20 ng of E. coli lysate containing 6-His-tagged protein. Cells were lysed with SDS sample buffer. The PVDF membrane was blocked with 5% dry milk in PBST for 1 h at RT. The primary antibody anti-6-His-tag mouse monoclonal, 6His was diluted in blocking buffer (antibody concentration 0.125 µg/ml) and incubated for 1 h at RT. The secondary antibody goat anti-mouse IgG polyclonal, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Western blot analysis of E. coli lysate containing 6-His-tagged protein with anti-6-His-tag antibody. Western blot analysis was performed on 50 ng or 25 ng of E. coli lysate containing 6-His-tagged protein. Cells were lysed with SDS sample buffer. The PVDF membrane was blocked with 5% dry milk in PBST for 1 h at RT. The primary antibody anti-6-His-tag mouse monoclonal, 6His was diluted in blocking buffer (antibody concentration 0.05 µg/ml) and incubated for 1 h at RT. The secondary antibody goat anti-mouse IgG polyclonal, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Key Features
Mouse monoclonal Suitable for ICC, IP and WB Reacts with 6-His-tag
Isotpye: IgG1
Product description
| Quantity | 100 µl |
|---|---|
| Antibody Type | Monoclonal |
| Host | Mouse |
| Isotype | IgG1 |
| Conjugate | Unconjugated |
| Application | ICC/IF, IP, WB |
| Purification | Affinity chromatography |
| Reactivity | 6-His |
| Storage | Short term at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles |
| Intended use | Research use only |
| Clone | 6His |
| Immunogen | Recombinant protein containing the sequence HHHHHH |
| Concentration | 250 µg/ml |
| Formulation | PBS, pH 7.4 with 0.09% sodium azide and 0.5% BSA |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunocytochemistry (ICC)/ Immunofluorescence (IF) | Assay dependent |
| Immunoprecipitation (IP) | Assay dependent |
| Western Blot (WB) | 1:2,000-1:5,000 (0.125-0.05 µg/ml) |
Background
The monoclonal 6-His antibody recognizes polyhistidine (6-His). The 6-His tag is commonly added to recombinant proteins and can be used for detection or purification of the tagged protein.
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FAQs
The concentration of unpurified supernatant, ascites, unpurified guinea pig serum and unpurified rabbit serum is not determined.
The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at support@progen.com.
The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at support@progen.com.
Most of our purified mouse antibodies contain 0.5% BSA as stabilizer. If BSA was added to the antibody solution, it is stated in the datasheet.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
Lyophilized antibodies can be stored at 2-8°C until expiration.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
The expiration date of our antibodies is indicated on the product label.
Most of our antibodies contain 0.09% sodium azide as preservative. If a preservative is added, it is mentioned in the datasheet.
The optimal antibody dilution for your specific protocol and application needs to be titrated in your lab with your equipment and sample. The optimal dilution may vary between protocols and samples. A good dilution for starting the titration is the dilution mentioned in the datasheet. If the sample needs a specific treatment (eg. Antigen retrieval for IHC on FFPE sections) this should also be mentioned on the antibody datasheet.
PROGEN antibodies are shipped at ambient temperature. The antibodies are stable at ambient temperature for the shipment period. Please store the antibodies as indicated in the datasheet upon arrival.
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