His-Tag Protein Expression Check Kit (ab270048)
Overview
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Product name
His-Tag Protein Expression Check Kit -
Product overview
His-Tag Protein Expression Check Kit (ab270048) is a quick competitive immunochromatography test that allows users to verify and monitor successful expression of His-tagged recombinant proteins before undertaking purification, as well as identifying fractions of most interest before performing other more labor intensive techniques such as Western blot or ELISA. The assay provides a qualitative yes/no answer, although a semi-quantitative measurement of the protein abundance in the sample can be determined using internal references and controls.
A His-tagged protein on the “Test line” (T line) and a ULFA-tagged protein on the “Control line” (C line) are immobilized on the nitrocellulose strips.
40nm Gold conjugated anti-HisTag antibody and 40 nm Gold conjugated anti-ULFA tag conjugates are dried on the conjugate pad and will selectively bind to the T and C line, respectively. The C line is assay-independent and should always appear as a strong red line; if it is not visible, the test is not valid and should be repeated.
His-Tag Protein Expression Check Kit is formulated as a competitive assay so that when the sample does not contain any His-tagged protein, the T line will appear as a strong red line. On the contrary, the T line will not be visible (or appear as a pale red line) if the sample contains the His- tagged protein that competes with the binding of the Gold conjugated anti-HisTag antibody conjugate to the antigen immobilized on the T line.
Benefits:
Quick and easy qualitative detection of His-tagged proteins
Compatible with cell culture media and lysate
No costly equipment required
Small sample volumes required (80 μL)
Compatible with Ni-NTA Affinity Resin - Amintra® (ab270549)
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Notes
This product is manufactured by Expedeon, an Abcam company. It was previously called His-Tag Check&Go!. Expedeon product code 4003-0030 is the same as the 30-test size of this product.
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 30 tests ab274087 - His-Tag Protein Expression Check Strips 1 x 30 tests ab274088 - Running Buffer (10X) 1 x 1500µl
Images
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How The Kit Works
His-Tag Protein Expression Check Kit (ab270048) is formulated as a competitive assay so that when the sample does not contain any His-tagged protein, the T line will appear as a strong red line (see Figure 1-A). On the contrary, if the sample does contain His-tagged protein, the T line will not be visible (or it will appear as a pale red line) as it competes with the binding of the Gold conjugated anti-HisTag antibody conjugate to the antigen immobilized on the T line (see Figure 1-B). The C line is assay-independent and should always appear as a strong red line – if it is not visible, the test is not valid and should be repeated.
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His-Tag Protein Expression Check KitSix different recombinant proteins were diluted down to 300, 100, 33 and 11ug/mL in 1X Running Buffer and run on a His-Tag Protein Expression Check strip. The strips were analysed using the EseQuant reader. Signal suppression is clearly detected in all the proteins tested.
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His-Tag Protein Expression Check KitNon induced (-I) and induced (+I) E. coli cell lysates expressing recombinant protein His-GST in 1X Running Buffer