Mouse Monoclonal 6X His tag® antibody - conjugated to Biotin. Suitable for ELISA, WB and reacts with Tag samples.
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 1% BSA, 0.88% Sodium chloride, 0.424% Potassium phosphate solution
ELISA | WB | |
---|---|---|
Tag | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Tag | Dilution info 1/200000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Tag | Dilution info 1/2000 - 1/10000 | Notes - |
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6 His epitope tag, HHHHHH epitope tag, HHHHHH tag, Hexa His tag, Polyhistidine Tag
Mouse Monoclonal 6X His tag® antibody - conjugated to Biotin. Suitable for ELISA, WB and reacts with Tag samples.
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 1% BSA, 0.88% Sodium chloride, 0.424% Potassium phosphate solution
This monoclonal anti-6X His tag antibody detects over-expressed proteins containing the 6X His epitope tag. To date, this antibody has reacted with all His tagged proteins so far tested. In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins. The antibody recognizes the His-tag (His-His-His-His-His-His) fused to either the amino- or carboxy-termini of targeted proteins in transfected or transformed cells.
Anti-6X HIS Epitope Tag was purified from concentrated tissue culture supernate by Protein A chromatography and is directed against the 6X His motif and is useful in determining its presence in various assays.
10-20 BAC molecules per Mouse Monoclonal Antibody
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The 6X His tag is a sequence commonly used for protein purification detection and immobilization. It consists of six histidine residues that bind to metal ions like nickel or cobalt. This makes it a highly efficient tag for affinity chromatography. His tag antibodies such as anti-His HRP facilitate the detection of His-tagged proteins in various assays. Being a small tag with a typical molecular weight of about 0.84 kDa it minimally interferes with protein function. Many proteins expressed in E. coli and other expression systems carry the 6X His tag for ease of purification and analysis.
Proteins tagged with the 6X His tag often play roles in studies involving recombinant proteins. The tag itself does not alter the biological function of the target protein. It is not part of any natural protein complex but aids in the study of such complexes. Researchers use His tag antibodies like anti-His to specifically target and study these tagged proteins without cross-reactivity to other proteins.
The incorporation of the 6X His tag into proteins allows them to be studied more efficiently in various biological pathways. It can be part of pathways like signal transduction or metabolic pathways. When tagged proteins interact with other molecules such as kinases or receptors researchers can track these interactions using the tag and anti-Histidine antibodies. This technique informs on proteins that are hard to purify due to low abundance or instability.
The 6X His tag itself does not have direct implications. However proteins of interest in disease models often carry the tag which allows researchers to purify and study them in detail. For example cancer-related proteins may be expressed with a His tag to understand their role in tumorigenesis. Similarly proteins involved in metabolic disorders could be tagged which helps in analyzing specific interactions or functions disrupted in diseases.
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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Block: 5% non-fat powdered milk for 1 hour at room temperature.
All lanes: Western blot - Biotin Anti-6X His tag® antibody [33D10.D2] (ab106261) at 1/5000 dilution
Lane 1: Purified histidine-tagged recombinant protein at 0.1 µg
Lane 2: E. coli cell lysate containing histidine-tagged expression construct at 0.2 µg
Lane 3: Purified GST-tagged recombinant protein at 0.1 µg
Lane 4: Purified FLAG-tagged recombinant protein at 0.1 µg
All lanes: Peroxidase mouse secondary antibody for 30 minutes at room temperature at 1/20000 dilution
Predicted band size: 1 kDa
ELISA analysis of ab106261 Mouse monoclonal [33D10.D2] to 6X His tag® (Biotin).
Antigen: HIS-tagged purified protein and E. coli cell lysates expressing HIS-Tagged construct, GST- and RON-tagged purified proteins.
Coating amount: 0.15 µg per welll.
Primary antibody: ab106261 at 100 µg/mL.
Dilution series: 2-fold.
Mid-point concentration: 200 ng/mL.
Secondary antibody: Peroxidase conjugated mouse secondary antibody at 1/10000.
Substrate: TMB ELISA Substrate.
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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