FACT (cdc68, Facilitates Chromatin Transcription)

FACT complex is a general chromatin factor that acts to reorganize nucleosomes. The FACT complex is involved in multiple processes that require DNA as a template such as mRNA elongation, DNA replication and DNA repair. During transcription elongation the FACT complex acts as a histone chaperone that both destabilizes and restores nucleosomal structure. It facilitates the passage of RNA polymerase II and transcription by promoting the dissociation of one histone H2A-H2B dimer from the nucleosome, then subsequently promotes the reestablishment of the nucleosome following the passage of RNA polymerase II. \n\nApplications: \nSuitable for use in Western Blot and Immunoprecipitation. Other applications not tested.\n\nRecommended Dilutions:\nWestern Blot: 1:1000-1:2000 dilution detects FACT/cdc68 in HeLa nuclear extracts. HeLa nuclear extract lysate was resolved by electrophoresis, transferred to PVDF and probed with F0004. Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP.\nImmunoprecipitation: Reported to immunoprecipitate FACT from HeLa nuclear extracts.\nOptimal dilutions to be determined by the researcher.\n\nIncluded Positive Antigen Control:\nH1838: HeLa Nuclear Extract. Add an equal volume of 2X Laemmli reducing sample buffer to 5-10ul of extract and boil for 5 min to reduce the preparation. Load 2-5ug of reduced lysate per lane for minigels.\n\nRecommended Secondary Antibodies:\nI1904-39: IgG, X-Adsorbed (HRP) Pab Gt xRb\nI1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt xRb\nI1904-46Q: IgG, H&L (HRP) Pab Gt xRb\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

F0004

Size

100ul

Applications

IP, WB

Hosts

Rabbit

Reactivities

Hum, Mouse

Form

Supplied as a liquid in 0.2M Tris, pH 8.0, 0.15M sodium chloride, 0.1mM EDTA, 5mg/ml BSA, 0.05% sodium azide, before the addition of glycerol to 30%.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1. Orphanides, G., et al., Nature 40: 284-288 (1999). 2. Yarnell, A.T., et al., J. Biol. Chem. 276: 25,736-25,741 (2001).

Additional Info

Recognizes mouse FACT/cdc68, Mr 140kD. A non-speci?c band is detected at ~120kD in longer exposures or when more that 5ug of nuclear extract is loaded per lane. Species Crossreactivity: human