PA1-862

antibody from Invitrogen Antibodies

Targeting: HDAC3 HD3, KDAC3, RPD3, RPD3-2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-862

Provider

Invitrogen Antibodies

Product name

HDAC3 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA1-862 detects histone deacetylase 3 (HDAC3) from canine, hamster, human and rat tissues and cells.

Concentration

1 mg/mL

References

Submitted references

Regulated clearance of histone deacetylase 3 protects independent formation of nuclear receptor corepressor complexes.

Guo C, Gow CH, Li Y, Gardner A, Khan S, Zhang J
The Journal of biological chemistry 2012 Apr 6;287(15):12111-20

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The transcriptional repressor Sp3 is associated with CK2-phosphorylated histone deacetylase 2.

Sun JM, Chen HY, Moniwa M, Litchfield DW, Seto E, Davie JR
The Journal of biological chemistry 2002 Sep 27;277(39):35783-6

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Effect of estradiol on histone acetylation dynamics in human breast cancer cells.

Sun JM, Chen HY, Davie JR
The Journal of biological chemistry 2001 Dec 28;276(52):49435-42

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Characterization of a human RPD3 ortholog, HDAC3.

Emiliani S, Fischle W, Van Lint C, Al-Abed Y, Verdin E
Proceedings of the National Academy of Sciences of the United States of America 1998 Mar 17;95(6):2795-800

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Differential display cloning of a novel human histone deacetylase (HDAC3) cDNA from PHA-activated immune cells.

Dangond F, Hafler DA, Tong JK, Randall J, Kojima R, Utku N, Gullans SR
Biochemical and biophysical research communications 1998 Jan 26;242(3):648-52

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of HDAC3 from HeLa cell lysate using Product # PA1-862.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of HDAC3 in HeLa Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a HDAC3 polyclonal antibody (Product # PA1-862) at a dilution of 1:200 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552). HDAC3 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry was performed on normal biopsies of deparaffinized human brain tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a Rabbit Polyclonal Antibody recognizing HDAC3 (Product # PA1-862) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin immunoprecipitation analysis of HDAC3 was performed using cross-linked chromatin from 1x10^6 HCT116 colon carcinoma cells treated with serum for 0, 15, and 30 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of an HDAC3 polyclonal antibody (Product # PA1-862). Chromatin aliquots from ~1x10^5 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify -15kb upstream of the Egr1 gene or exon-1 or exon-2 of Egr1. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the Egr-1 locus is shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by Egr-1 primers are represented by black bars. Data courtesy of the Innovators Program.