Human/Mouse/Rat RBBP4 Antibody

  
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human RBBP-4 in direct ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG2B Clone # 968701
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human RBBP-4
    Ala2-Ser425
    Accession # 09028
  • 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
    0.5 µg/mL
    See below
  • Immunohistochemistry
    5 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, Mouse, and Rat RBBP4 by Western Blot. Western blot shows lysates of K562 human chronic myelogenous leukemia cell line, Neuro‑2A mouse neuroblastoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and
Rat‑2 rat embryonic fibroblast cell line. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat RBBP4 Monoclonal Antibody (Catalog # MAB7416) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for RBBP4 at approximately 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
RBBP4 in Human Breast Cancer Tissue.

Retinoblastoma Binding Protein 4 (RBBP4) was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human/Mouse/Rat RBBP4 Monoclonal Antibody (Catalog # MAB7416) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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: RBBP4

RBBP4 (Retinoblastoma-Binding Protein 4; also RbAp48 and CAF-1) is a 48-56 kDa member of the WD Repeat RBAP46/48/MSI1 family of proteins. It is ubiquitously expressed, and acts as a transcriptional repressor. At the start of G1 of the cell cycle, Rb (retinoblastoma protein) normally associates with an E2F transcription complex on E2F responsive genes, blocking E2F activity. At the appropriate time, Rb is phosphorylated, causing its dissociation from E2F and resulting in E2F activation. Dephosphorylated Rb apparently mediates transcriptional repression by recruiting a histone deacetylase (HDAC), followed by HDAC binding to RBBP4. RBBP4, being a histone-binding protein, now brings histones plus HDACs together, resulting in histone deacetylation and gene silencing. Human RBBP4 is 425 amino acids (aa) in length. It contains six WD repeats (aa 122-403), two N-terminal acetylation sites, and two serine phosphorylation sites at positions 110 and 146. There are at least two isoform variants. One contains a six aa substitution for aa 405-425, while another utilizes an alternative start site at Met36. Human and mouse RBBP4 are identical in aa sequence.

  • Long Name:
    Retinoblastoma Binding Protein 4
  • Entrez Gene IDs:
    5928 (Human); 19646 (Mouse); 685491 (Rat)
  • Alternate Names:
    CAF-1 p48; CAF-1 subunit C; CAF-1C; CAF-I p48; Chromatin assembly factor 1 subunit C; Chromatin assembly factor I p48 subunit; chromatin assembly factor/CAF-1 p48 subunit; histone-binding protein RBBP4; MSI1 protein homolog; Nucleosome-remodeling factor subunit RBAP48; NURF55; RBAP48; RBBP4; RBBP-4; retinoblastoma binding protein 4; Retinoblastoma-binding protein 4CAF-I 48 kDa subunit; Retinoblastoma-binding protein p48
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