Prolactin Antibody (1085531) [Unconjugated]

Western Blot shows lysates of human pituitary tissue. PVDF membrane was probed with 0.5 µg/ml of Mouse Anti-Human Prolactin Monoclonal Antibody (Catalog # MAB11590) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Prolactin at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Simple Western shows lysates of human pituitary tissue, loaded at 0.2 mg/ml. A specific band was detected for Prolactin at approximately 29 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human Prolactin Monoclonal Antibody (Catalog # MAB11590). This experiment was conducted under reducing conditions and using the 12‑230 kDa separation system.

Prolactin was detected in immersion fixed paraffin-embedded sections of human pituitary using Mouse Anti-Human Prolactin Monoclonal Antibody (Catalog # mab11590) at 5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC
• Clone: 1085531
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Prolactin Antibody (1085531) [Unconjugated] Summary

• Immunogen: E. coli-derived human Prolactin
  Leu29-Cys227
  Accession # P01236
• Specificity: Detects rhProlactin in Direct ELISA
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: PRL
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-25 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Prolactin Antibody (1085531) [Unconjugated]

• PRL
• Prolactin

Background

Prolactin (gene name PRL) is a secreted neuroendocrine pituitary hormone that acts primarily on the mammary gland to promote lactation, but has pleiotropic effects in both males and females (1-6). Prolactin is predominantly found as 199 amino acid, 25 kDa glycosylated and 23 kDa non-glycosylated monomers (6). Human prolactin shares only 60% and 63% amino acid sequence identity with mouse and rat prolactin, respectively, although rat prolactin can activate the human prolactin receptor (3). Post-translational modifications such as polymerization, complex formation with IgG (in humans), glycosylation, and proteolytic cleavage can alter the activities of prolactin (6-8). Non-glycosylated prolactin is produced by the pituitary and packaged in storage granules before secretion, while glycosylated prolactin is reported to be constitutively secreted, have lower biological potency, and be removed from the circulation more quickly (3, 6, 7). Cleavage by matrix metalloproteinases or Cathepsin D can produce N-terminal 16 kDa antiangiogenic fragments also called vasoinhibins (9, 10). Thrombin can produce C-terminal 16 kDa fragments that are not antiangiogenic (3). Prolactin is synthesized mainly by the anterior pituitary in all mammals, where secretion is under tonic inhibition by hypothalamic dopamine (2, 3). In humans, prolactin is also produced peripherally (2-5). Prolactin expression is low during early human pregnancy, but increases in late pregnancy (2, 3). The prolactin receptor (gene name PRLR) is a transmembrane type I glycoprotein that belongs to the cytokine hematopoietic receptor family. Expression of the prolactin receptor is widespread (2-5). Each prolactin molecule is thought to bind two receptor molecules (11). In addition to its lactogenic activity, peripherally produced prolactin plays roles in breast and prostate cancer development, regulation of reproductive function, and immunoregulation (5, 6).

1. Cooke, N.E. et al. (1981) J. Biol. Chem. 256:4007.
2. Grattan, D.R. and I.C. Kokay (2008) J. Neuroendocrinol. 20:752.
3. Ben-Jonathan, N. et al. (2008) Endocr. Rev. 29:1.
4. Bernichtein, S. et al. (2010) J. Endocrinol. 206:1.
5. Goffin, V. et al. (2011) Nat. Rev. Urology 8:597.
6. Price, A.E. et al. (1995) Endoc. 136:4827.
7. Hoffmann, T. et al. (1993) J. Endoc. Invest. 16:807.
8. Cole, E. et al. (1991) Endoc. 129:2639.
9. Piwnica, D. et al. (2006) Mol. Endocrinol. 20:3263.
10. Macotela, Y. et al. (2006) J. Cell Sci. 119:1790.
11. Broutin, I. et al. (2010) J. Biol. Chem. 285:8422.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: PRL
• Uniprot:
  • P01236()