Mouse CRISP-1 Antibody

  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse CRISP-1 in direct ELISAs.
  • Source
    Monoclonal Rat IgG2B Clone # 840017
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse CRISP-1
    Gln20-His244
    Accession # Q03401
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunohistochemistry
    8-25 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize CRISP‑1-inhibited proliferation in the 3A‑sub E human placenta cell line. The Neutralization Dose (ND50) is typically 10-20 µg/mL in the presence of 10 µg/mL Recombinant Mouse CRISP‑1.
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

Proliferation Inhibited by CRISP‑1 and Neutralization by Mouse CRISP‑1 Antibody. Recombinant Mouse CRISP‑1 inhibits proliferation in the 3A‑sub E human placenta cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Proliferation inhibited by Recombinant Mouse CRISP‑1 (10 µg/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Mouse CRISP‑1 Monoclonal Antibody (Catalog # MAB4675). The ND50 is typically 10-20 µg/mL.

Detection of Mouse CRISP‑1 by Western Blot. Western blot shows lysates of mouse epididymis tissue. PVDF membrane was probed with 1 µg/mL of Rat Anti-Mouse CRISP‑1 Monoclonal Antibody (Catalog # MAB4675) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for CRISP‑1 at approximately 27-30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
CRISP‑1 in Mouse Epididymis. CRISP‑1 was detected in perfusion fixed frozen sections of mouse epididymis using Rat Anti-Mouse CRISP‑1 Monoclonal Antibody (Catalog # MAB4675) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
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: CRISP-1
Cysteine-rich Secretory Protein 1 (CRISP‑1), also known as Acidic Epididymal Glycoprotein 1 (AEG1) and Sperm‑Coating Glycoprotein 1 (SCP1), is a 29-32 kDa (26.4 kDa predicted) protein that is involved in male reproductive biology (1, 2). CRISPs belong to the CAP superfamily of molecules that also includes several snake, insect, and lizard venom proteins (3). Structurally, CRISPs consist of an N-terminal SCP/CAP domain, a hinge region, and a Cys-rich domain with 16 invariant cysteine residues (4). CRISPs are expressed in discrete but overlapping regions of the male reproductive tract and play a role in spermatozoa adhesion with Sertoli cells and oocytes, decapacitation, and the acrosomal reaction (1). Expression patterns, genomic structure, and sequence conservation indicate that the likely ortholog of human CRISP-1 in mouse and rat is CRISP-4 (5, 6). Mature mouse CRISP-1 shares 68% amino acid (aa) sequence identity with rat CRISP-1. It shares 55%, 73%, and 43% aa sequence identity with mouse CRISP-2, -3, and -4, respectively. Rodent CRISP-1 is secreted by the epididymal epithelium and associates with the surface of spermatozoa in the epididymis where it inhibits capacitation (7-9). It is released from the sperm cell surface once capacitation begins in the female reproductive tract or in vitro (10). CRISP-1 is involved in the interaction of spermatozoa with the oocyte zona pellucida as well as in the fusion of sperm and egg (11, 12). CRISP-1 is additionally expressed in the lower medulla of hair shafts and under androgen control in the submandibular salivary glands of male mice (13-15).
  • References:
    1. Koppers, A.J. et al. (2011) Asian J. Androl. 13:111.
    2. Gibbs, G.M. et al. (2008) Endocr. Rev. 29:865.
    3. Yamazaki, Y. and T. Morita (2004) Toxicon 44:227.
    4. Guo, M. et al. (2005) J. Biol. Chem. 280:12405.
    5. Jalkanen, J. et al. (2005) Biol. Reprod. 72:1268.
    6. Nolan, M.A. et al. (2006) Biol. Reprod. 74:984.
    7. Charest, N.J. et al. (1989) Endocrinology 125:942.
    8. Maldera, J.A. et al. (2011) Biol. Reprod. 85:503.
    9. Nixon, B. et al. (2006) Biol. Reprod. 74:275.
    10. Roberts, K.P. et al. (2003) Biol. Reprod. 69:572.
    11. Busso, D. et al. (2007) Biol. Reprod. 77:848.
    12. Da Ros, V.G. et al. (2008) Dev. Biol. 320:12.
    13. Haendler, B. et al. (1993) Endocrinology 133:192.
    14. Mizuki, N. and M. Kasahara (1992) Mol. Cell. Endocrinol. 89:25.
    15. Peterson, R.L. et al. (2005) J. Investig. Dermatol. Symp. Proc. 10:238.
  • Long Name:
    Cysteine-rich Secretory Protein 1
  • Entrez Gene IDs:
    167 (Human); 11571 (Mouse); 64827 (Rat)
  • Alternate Names:
    Acidic epididymal glycoprotein homolog; acidic epididymal glycoprotein-like 1; AEGL1; AEG-related protein; ARPAEG-like protein; CRISP1; CRISP-1; CRISP-1AEGL1HSCRISP1D; cysteine-rich secretory protein 1; cysteine-rich secretory protein-1 delta; HSCRISP1G; HUMARP
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