This product was prepared from monospecific antiserum by a multi-step procedure which includes delipidation, salt fractionation and ion exchange chromatography. A single precipitin arc was observed against anti-Rabbit Serum when assayed by immunoelectrophoresis. Reacts with Human and Mouse. Does not recognise the A subunit.
Purification
Prepared from monospecific antiserum by delipidation, salt fractionation
Immunogen
NF-Y (B subunit specific) peptide corresponding to a region near the N-terminus of the Human protein conjugated to Keyhole Limpet Hemocyanin (KLH).
Suitable for Immunoprecipitation, Western blotting (1/500), ELISA (1/4,000) and supershiftassays. Recommended Dilutions: This product was assayed by immunoblot and found to bereactive against the 25 kDa B subunit of NF-Y at a dilution of 1: 500 followed by reactionwith Peroxidase conjugated Affinity Purified anti-Rabbit IgG [H&L] (Goat). Anti-NF-Y (Bsubunit specific) is suitable for the detection by immunoblot of human and mouse NF-Y (Bsubunit specific). Minimal reaction was observed by immunoblot against the 35 kDa Asubunit of NF-Y. This product was also tested in a gel supershift assay and found to bereactive against human and mouse NF-Y using 2.0 to 4.0 l per assay. This product was assayed against NF-Y B subunit peptide in an antibody capture ELISAusing Peroxidase conjugated Affinity Purified anti-Rabbit IgG [H&L] (Goat) and ABTSsubstrate. A dilution of 1: 4000 is suggested from this experiment. Minimal reactivity was
Restrictions
For Research Use only
Concentration
1.0 mg/mL (by UV absorbance at 280 nm)
Buffer
0.01 M Sodium Phosphate, 0.11 M Sodium Chloride, pH 7.2 with 0.01 % Sodium Azide as preservative.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing. Dilute only to immedoate use.
Storage
-20 °C
Storage Comment
Store original vial at -20 °C or below. After opening aliquot contents and freeze at -20 °C or below.
Target
NFYB
(Nuclear Transcription Factor Y, beta (NFYB))
The Y box is a CCAAT box which is bound by the heteromeric DNA binding protein, NF-Y (also known as CBF and CP1). Unlike the transcription factors C/EBP and CTF/NF1 which also bind CCAAT-like sequences, NF-Y exhibits a strict binding requirement for this pentanucleotide sequence. Binding sites for this factor have been described for nearly 30 % of all eukaryotic genes. Y/CCAAT sequences were frequently observed in the promoter-proximal sequences. NF-Y is composed of 3 separate subunits (A,B and C) each of which is required for DNA binding. Each subunit has remained highly conserved throughout evolution. In fact, homologous yeast subunits can substitute for mammalian NF-Y in DNA-binding assays. The conserved core sequences of NF-YB and NF-YC contain a 70 aa region similar to the histone fold motif of nucleosomes H2A and H2B. The unique structure and evolutionary conservation of this transcription factor suggests that it plays a fundamental role in the readout of eukaryotic genetic information.Synonyms: HAP3, Nuclear transcription factor Y subunit beta