Human Lysine (K)-specific Demethylase 5B/KDM5B Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human KDM5B in Western blots.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human KDM5B
    Leu819-Arg918
    Accession # Q9UGL1
  • 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
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Lysine (K)-specific Demethylase 5B/KDM5B by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line. PVDF Membrane was probed with 1 µg/mL of Human Lysine (K)-specific Demethylase 5B/KDM5B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6477) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Lysine (K)-specific Demethylase 5B/KDM5B at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Lysine (K)-specific Demethylase 5B/KDM5B in BT‑20 Human Cell Line. Lysine (K)-specific Demethylase 5B/KDM5B was detected in immersion fixed BT‑20 human breast cancer cell line using Human Lysine (K)-specific Demethylase 5B/KDM5B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6477) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • 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: Lysine (K)-specific Demethylase 5B/KDM5B

KDM5B (also JARID1B, RBP2-H1 and PLU-1) is a 170-200 kDa member of the JARID1 histone demethylase family of proteins. It is a transcriptional repressor that is highly expressed in Sertoli cells and mammary epithelium during pregnancy, and interacts with BF-1 and PAX9. KDM5B is reported to demethylate tri- and di-methylated Lys4 on histone H3. It has no activity on Lys9 or 27. Human KDM5B is 1544 amino acids (aa) in length. It contains a jumonji N domain (aa 32-73), an ARID DNA binding region (aa 97-187), a PHD domain (aa 309-359), a jumonji C domain (aa 486-602), one C5HC2 motif (aa 692-745), and two consecutive PHD domains (aa 1176-1538). There are three utilized phosphorylation sites at Ser1383, 1384 and 1405.There are three potential splice variants. Two show a premature truncation after Pro1433 coupled to an alternative start site at either Met396 or Met159. A third contains a 36 aa insertion after Glu237. Over aa 819-918, human KDM5B shares 96% aa identity with mouse KDM5B.

  • Entrez Gene IDs:
    10765 (Human); 75605 (Mouse); 304809 (Rat)
  • Alternate Names:
    CT31; CT31FLJ10538; EC 1.14.11; EC 1.14.11.-; homolog 1A; Jarid1B; JARID1BFLJ16281; Jumonji, AT rich interactive domain 1B (RBP2-like); jumonji, AT rich interactive domain 1B; KDM5B; Lysine (K)specific Demethylase 5B; Lysine (K)-specific Demethylase 5B; PLU-1; PLU-1FLJ12459; PLU1FLJ23670; PUT1; RBBP2H1; RBBP2H1AFLJ12491; RBP2-H1; Retinoblastoma-binding protein 2 homolog 1
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