詳細(xì)說明
Species Reactivity
Human, Mouse
Specificity
Detects human HDAC8 in direct ELISAs and Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human HDAC8 isoform 1
Met1-Val377 (Leu31Pro)
Accession # Q9BY41Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 μm filtered solution in PBS.
Label
Unconjugated
Applications
Recommended
ConcentrationSample
Western Blot
1 μg/mL
See below
Chromatin Immunoprecipitation (ChIP)
5 μg/5 x 10 6 cells
See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. are available in the Technical Information section on our website.
Data Examples
Western Blot | 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 # ). 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 . |
Chromatin Immunoprecipitation (ChIP) | 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 # ) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Sheep IgG Secondary Antibody (Catalog # Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # ) and DNA was purified using chelating resin solution. The IFN-gamma promoter was detected by standard PCR. |
Preparation and Storage
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.
Background: Histone Deacetylase 8/HDAC8
HDAC8 (Histone deacetylase 8) is a 50 kDa member of the class I subfamily, histone deacetylase family of transcriptional regulators. It is a smooth muscle enzyme that deacetylates histone lysines, blocking gene transcription. HDAC8 appears to function as a monomer, and phosphorylation at Ser39 inhibits activity. Human HDAC8 is 377 amino acids (aa) in length. It contains a series of alpha helices and strands, plus a catalytic ferrous ion binding site (Asp178-His180-Asp267). There are four potential splice variants, all of which possess an alternate start site 38 aa upstream of the standard start site. Three have additional changes; two show a one and three aa substitution respectively for aa 211-377, while a third has a 15 aa substitution for aa 148-377. Human HDAC8 shares more than 96% aa identity with both mouse and canine HDAC8.
Entrez Gene IDs:
55869 (Human); 70315 (Mouse); 363481 (Rat)
Alternate Names:
EC 3.5.1.98; HD8; HDAC8; HDACL1; Histone Deacetylase 8; histone deacetylase-like 1; RPD3