Histone Deacetylase 2 (HDAC2, Tanscriptional Regulator Homolog RPD3, HD2)

Applications: \nSuitable for use in Western Blot, Immunoprecipitation and Immunocytochemistry. Other applications not tested.\n\nRecommended Dilution:\nImmunocytochemistry: Reported by an independent laboratory to show positive immunostaining for HDAC2 in NIH/3T3 and U2OS cells fixed with paraformaldehyde.\n\nWestern Blot: 0.5-2ug/ml Detects HDAC2 in nuclear extracts from HeLa cells. HeLa nuclear extract was resolved by electrophoresis, transferred to nitrocellulose and probed with 0.5ug/ml H5109-47B. Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system. \n\nImmunoprecipitation and HDAC Assay: 3ul was used to immunoprecipitate HDAC activity from HeLa nuclear extract, which was then measured using H5109-62 (HDAC Assay Kit; Fluorometric Detection). An independent laboratory has also demonstrated positive immunoprecipitation/HDAC assay results using NIH/3T3 cells. \nOptimal dilutions to be determined by the researcher.\n\nPositive Control: \nH1838 HeLa cell extract:\nAdd an equal volume of Laemmli reducing sample buffer to 10ul of lysate. Boil for 5 minutes to reduce the preparation. Load 10-20ug of reduced lysate per lane for Western Blot.\n\nRecommended secondary antibodies:\nI1904-06C: IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo\nI1904-06H IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

H5109-47B

Size

200ug

Applications

ICC, IP, WB

Hosts

Mouse

Reactivities

Hum, Mouse, Bov

Form

Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, before the addition of glycerol to 30%

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG1

References

1. Ahringer, J., Trends Genet. 16:351-356, 2000. 2. Humphrey, G.W., et al., J. Biol. Chem. 276:6817-6824, 2001. 3. You, A., et al., Proc. Natl. Acad. Sci. USA. 98:1454-1458, 2001. 4. Yang, W.M., et al., Proc. Natl. Acad. Sci. USA. 93:12845-12850, 1996.

Additional Info

Recognizes human HDAC2, Mr 60kD. Species Crossreactivity: mouse, bovine and hamster. Species sequence homology: rat.