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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Chromatin Immunoprecipitation [1]
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- Product number
- AF4359 - Provider product page
- Provider
- R&D Systems
- Product name
- Human/Mouse Histone Deacetylase 8/HDAC8 Antibody
- Antibody type
- Polyclonal
- Description
- Immunogen affinity purified. Detects human HDAC8 in direct ELISAs and Western blots.
- Reactivity
- Human, Mouse
- Host
- Sheep
- Conjugate
- Unconjugated
- Antigen sequence
Q9BY41- Isotype
- IgG
- Vial size
- 100 ug
- Concentration
- LYOPH
- Storage
- Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.
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Supportive validation
- Submitted by
- R&D Systems (provider)
- Main image
- Experimental details
- Detection of Human Histone Deacetylase 8/HDAC8 by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse Histone Deacetylase 8/HDAC8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4359) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Histone Deacetylase 8/HDAC8 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Supportive validation
- Submitted by
- R&D Systems (provider)
- Main image
- Experimental details
- Detection of Histone Deacetylase 8/ HDAC8-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Histone Deacetylase 8/HDAC8/DNA complexes were immunoprecipitated using 5 μg Sheep Anti-Human/Mouse Histone Deacetylase 8/HDAC8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4359) or control antibody (Catalog # 5-001-A) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Sheep IgG Secondary Antibody (Catalog # BAF016). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The IFN-gamma promoter was detected by standard PCR.
