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Human/Mouse Histone Deacetylase 8 Affinity Purified PAb 100 UG

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Human/Mouse Histone Deacetylase 8 Affinity Purified PAb 100 UG信息二維碼

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產(chǎn)品介紹

    基本參數(shù)

    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 # Q9BY41

    • Formulation

      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    
      Concentration

      Sample

    • 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







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