Description
NF-κB is a transcription factor that is a member of the mammalian NF-κB/Rel family of proteins. Members of this family are involved in the regulation of cell proliferation, immune function, as well as development. In resting cells, IκBα binds to and maintains NF-κB in the cytoplasm by blocking the nuclear localization sequences of NF-κB. In the cellular response to an extracellular signal, IκBα is phosphorylated and subsequently degraded via the ubiquination-proteasome pathway, allowing NF-κB to translocate to the nucleus. Once in the nucleus, NF-κB can induce the transcription of IκBα thereby renewing the cycle so that IκBα can form a complex with NF-κB and maintain it in its cytoplasmic location. IκBα -/- mice show an increased level of NF-κB activity and have been shown to die soon after birth. The 25/IkBa/MAD-3 monoclonal antibody recognizes human IκBα regardless of phosphorylation status and does not cross-react with mouse IκBα.
This Alexa Fluor® 647-conjugated antibody is suitable for intracellular staining of cell lines and primary cells using BD Cytofix™ Fixation Buffer or BD Phosflow™ Lyse/Fix Buffer. Although this antibody can be used with BD Phosflow™ Perm/Wash Buffer I or BD Phosflow™ Perm Buffers II, III or IV, it performs optimally when used with BD Phosflow™ Perm Buffer II.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
7. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
9. An isotype control should be used at the same concentration as the antibody of interest.
