Alexa Fluor® 647 Mouse anti-IRF-7 (pS477/pS479)
Clone K47-671 (RUO)
- Brand BD Phosflow™
- Vol. Per Test 20 µl
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Human (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated Human IRF-7 Peptide
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
Interferon regulatory factor 7 (IRF-7) is a transcription factor that regulates anti-viral defenses by controlling the induction of type-I interferon (IFN) responses. IRF-7 expression is induced in lymphoid cells by virus infection, as well as by IFN, lipopolysaccharide, and TNF-α. IRF-7 responses are initiated by Toll-like receptors (TLR) or the cytoplasmic protein retinoic acid inducible gene I (RIG-I). Upon TLR activation, it forms cytoplasmic complexes with MyD88, an adaptor in the TLR signaling pathways. The TLR-dependent and RIG-I-dependent pathways activate kinases, such as IKK-ε and TBK1, that phosphorylate IRF-7 and induce movement of IRF-7-containing complexes to the nucleus, where it preferentially activates IFN-α promoters.
The K47-671 monoclonal antibody recognizes human IRF-7 phosphorylated at serines 477 and 479 (pS477/pS479). Our in-house testing is performed on a cell line that has been co-transfected with TBK1 and IRF-7. Phosphorylation of IRF-7 in the transfectants requires virus infection or over-expression of a signaling molecule of the RIG-I pathway, such as TBK1. Phosphorylation of endogenous IRF-7 in untransfected cells has not yet been detected. We confirmed that mAb K47-671 does not cross-react with TBK1 by Western blot analysis using the purified antibody.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
- 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).
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.