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Nushift™ AP-1 Family

Catalog No: 37003

Format: 17 rxns

Reactivity:

Human, Mouse

View Data Sheet

$ 995

Background:

This kit is designed for the detection of c-Fos, c-Jun, FosB, JunB and JunD using the electrophoretic mobility shift assay (EMSA). Applications include screening for the presence of c-Fos, c-Jun, FosB, JunB and JunD in various protein sources, evaluating the functional binding of c-Fos, c-Jun, FosB, JunB and JunD recombinant proteins, analysis of different DNA-binding consensus sites and characterization of promoter elements.

The following are sufficient for 17 supershift reactions:
2 x 40 µl AP-1 Oligo WT
12 µl AP-1 Oligo probe WT
2 x 40 µl AP-1 mutant Oligo
6 x 40 µl 3T3 SR nuclear extract (c-Fos, FosB, JunB detection)
100 µl c-Fos rabbit pAb
100 µl c-Jun rabbit pAb
100 µl FosB rabbit pAb
17 µl JunB rabbit pAb
100 µl JunD rabbit pAb
2 x 40 µl 3T6 nuclear extract (c-Jun detection)
2 x 40 µl Jurkat nuclear extract (TPA + CI stimulated) (JunD detection)
30 µl each c-Fos, c-Jun, FosB, JunB, JunD Peptide
2 x 240 µl Dilution Buffer A
3 x 240 µl Binding Buffer B-1
2 x 240 µl Binding Buffer C-1
2 x 360 µl Stabilizing Solution D
5 MicroSpin™ G-25 Columns

Application Notes:

When performing EMSA experiments individual optimization of extract and antigen quantities may be required. We recommend the following conditions when using the AP-1 Family Kit. For c-Fos use 9 µg of extract and 2-10 µl of antibody. For c-Jun use 15 µg of extract and 6 µl of antibody. For FosB use 9 µg of extract and 4 µl of antibody. For JunB use 9 µg of extract and 2 µl of antibody. For JunD use 15 µg of extract and 6-10 µl of antibody. We also recommend the use of 0.25X Tris-boric acid electrophoresis buffer.

Storage:

Upon receipt, the nuclear extract(s) must be stored at -80°C and the Microspin&trade columns should be stored at room temperature. All other components may be stored at either -20°C or -80°C. We recommend aliquoting the extracts into single-use fractions and then storing them at -80°C. This avoids repeated freeze/thaw cycles.

Guarantee:

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

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Nushift™ AP-1 Family

Background:

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Application Notes:

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Guarantee: