Human/Mouse/Rat Rab27a Antibody

  
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat RAB27A in Western blots and detects recombinant human RAB27A in direct ELISAs. In direct ELISAs, less than 1% cross-reactivity with recombinant human RAB27B is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human RAB27A
    Ser135-Ala218
    Accession # P51159
  • 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
  • Simple Western
    5 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, Mouse, and Rat RAB27A by Western Blot. Western blot shows lysates of K562 human chronic myelogenous leukemia cell line, 786‑O human renal cell adenocarcinoma cell line, MCF‑7 human breast cancer cell line, Jurkat human acute T cell leukemia cell line, Rat‑2 rat embryonic fibroblast cell line, and Neuro‑2A mouse neuroblastoma cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human/Mouse/Rat RAB27A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7245) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for RAB27A at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
RAB27A in U937 Human Cell Line. RAB27A was detected in immersion fixed U937 human histiocytic lymphoma cell line using Sheep Anti‑Human/Mouse/Rat RAB27A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7245) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Human Rab27a by Simple WesternTM. Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Rab27a at approximately 32 kDa (as indicated) using 5 µg/mL of Sheep Anti-Human/Mouse/Rat Rab27a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7245) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
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: Rab27a

RAB27A (Ras-related protein Rab 27A; also GTP-binding protein Ram) is a 27-28 kDa member of the Rab27 subfamily, Rab family, Small GTPase superfamily of proteins. It is widely expressed, and found in cells diverse as mast cells, cytotoxic T cells, melanocytes, retinal pigment epithelium and pancreatic beta -cells. RAB27A plays a key role in the secretion of specialized lysosomes termed secretory lysosomes. In melanocytes, for example, RAB27A is incorporated into the melanosome membrane where it serves as a docking factor for melanophilin and myosin-Va, regulating melanosome transport to, and concentration at, sites of release. Human RAB27A is 221 amino acids (aa) in length. It contains multiple Rab family and subfamily motifs, and concludes with a C-terminal CXC prenylation sequence (aa 219‑221). There is one potential splice variant that shows a deletion of aa 146-153. Over aa 135-218, human RAB27A shares 92% and 94% aa sequence identity with mouse Rab27A and rat RAB27A, respectively.

  • Long Name:
    RAs Genes from Brain Protein 27A
  • Entrez Gene IDs:
    5873 (Human)
  • Alternate Names:
    GS2; GS2rab-27; GTP-binding protein Ram; HsT18676; Rab-27; Rab27a; RAB27A, member RAS oncogene family; RAB27MGC117246; RAM; RAMras-related protein Rab-27A
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