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Human Prolactin Antibody

Catalog #: AF682
6 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF682
AF682-SP
Detection of Human Prolactin by Western Blot.
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Citations (6)
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Human Prolactin Antibody Summary

Species Reactivity
Human
Specificity
Detects human Prolactin in ELISAs and Western blots. In sandwich ELISAs, less than 0.05% cross-reactivity with recombinant mouse Prolactin and recombinant human Prolactin R.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human Prolactin
Leu29-Cys227
Accession # Q5THQ0
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or 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
0.25 µg/mL
See below
Simple Western
2.5 µg/mL
Human pituitary tissue
Immunohistochemistry
1-15 µg/mL
See below

Human Prolactin Sandwich Immunoassay

Recommended Concentration
Reagent
ELISA Capture (Matched Antibody Pair)
0.2-0.8 µg/mL 

Use in combination with:

Detection Reagent: Human Prolactin Biotinylated Antibody (Catalog # BAF682)

Standard: Recombinant Human Prolactin Protein, CF (Catalog # 682-PL)

Neutralization
Measured by its ability to neutralize Prolactin-induced proliferation in the Nb2‑11 rat lymphoma cell line [Gout, P.W. et al. (1980) Cancer Research 40:2433]. The Neutralization Dose (ND50) is typically 0.02-0.05 µg/mL in the presence of 0.5 ng/mL Recombinant Human Prolactin.

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.

Scientific Data

Western Blot Detection of Human Prolactin antibody by Western Blot. View Larger

Detection of Human Prolactin by Western Blot. Western blot shows lysates of human pituitary tissue. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF682) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Prolactin at approximately 23 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry Prolactin antibody in Human Testis by Immunohistochemistry (IHC-P). View Larger

Prolactin in Human Testis. Prolactin was detected in immersion fixed paraffin-embedded sections of human testis using Goat Anti-Human Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF682) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of sperm cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Neutralization Cell Proliferation Induced by Prolactin and Neutralization by Human Prolactin Antibody. View Larger

Cell Proliferation Induced by Prolactin and Neutralization by Human Prolactin Antibody. Recombinant Human Prolactin (Catalog # 682-PL) stimulates proliferation in the Nb2‑11 rat lymphoma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human Prolactin (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF682). The ND50 is typically 0.02-0.05 µg/mL.

Simple Western View Larger

Detection of Human Prolactin by Simple WesternTM. Simple Western shows lysates of human pituitary, loaded at 0.2 mg/ml. A specific band was detected for Prolactin at approximately 30 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Human Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF682). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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: Prolactin

Prolactin (PRL) is a neuroendocrine pituitary hormone. Prolactin is synthesized by the anterior pituitary, placenta, brain, uterus, dermal fibroblasts, decidua, B cell, T cells, NK cells, and breast cancer cells. Originally characterized as a lactogenic hormone, studies have demonstrated broader roles in breast cancer development, regulation of reproductive function, and immunoregulation. In the immune system, prolactin has been shown to be secreted by human PBMC and to act as a proliferative growth factor. Additionally, prolactin treatment of human PBMC has been shown to enhance IFN-gamma production. Prolactin has several molecular forms. The predominant form is a monomer, the non-glycosylated form is 23 kDa and the glycosylated form is 25 kDa. Glycosylated prolactin is removed from the circulation faster and has been reported to have lower biological potency. Prolactin cDNA encodes a 227 amino acid residue protein with a putative 28 aa residue signal peptide. The prolactin receptor is a transmembrane type I glycoprotein that belongs to the cytokine hematopoietic receptor family. B cells, T cells, macrophages, NK cells, monocytes, CD34+ progenitor cells, neutrophils, mammary gland, liver, kidney, adrenals, ovaries, testis, prostrate, seminal vesicles, and hypothalamus have all been shown to express the prolactin receptor. Three forms of the receptor, generated by differential splicing, have been identified. These isoforms differ in the length of their cytoplasmic domains. It is believed that the short cytoplasmic form is non-functional. Prolactin signal transduction involves the JAK/STAT families and Src kinase family.

References
  1. Cooke, N.E. et al. (1981) J. Biol. Chem. 256:4007.
  2. Ben-Johnson, N. et al. (1996) Endoc. Rev. 17:639.
  3. Cesario, T. et al. (1994) Proc. Soc. Exp. Biol. Med. 205:89.
  4. Price, A.E. et al. (1995) Endoc. 136:4827.
  5. Hoffmann, T. et al. (1993) J. Endoc. Invest. 16:807.
  6. Bellone, G. et al. (1995) J. Cell Physiol. 163:221.
  7. Cole, E. et al. (1991) Endoc. 129:2639.
  8. Lewis, U. et al. (1985) Endoc. 116:359.
Entrez Gene IDs
5617 (Human); 19109 (Mouse)
Alternate Names
PRL; Prolactin

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Citations for Human Prolactin Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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  1. The role of Interleukin-18 in recurrent early pregnancy loss
    Authors: S Löb, B Ochmann, Z Ma, T Vilsmaier, C Kuhn, E Schmoeckel, SL Herbert, T Kolben, A Wöckel, S Mahner, U Jeschke
    Journal of reproductive immunology, 2021-10-05;148(0):103432.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  2. Interleukin-1 beta is significantly upregulated in the decidua of spontaneous and recurrent miscarriage placentas
    Authors: S Löb, N Amann, C Kuhn, E Schmoeckel, A Wöckel, A Zati Zehni, T Kaltofen, S Keckstein, JN Mumm, S Meister, T Kolben, S Mahner, U Jeschke, T Vilsmaier
    Journal of reproductive immunology, 2021-01-30;144(0):103283.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  3. Short prolactin isoforms are expressed in photoreceptors of canine retinas undergoing retinal degeneration
    Authors: R Sudharsan, L Murgiano, HY Tang, TW Olsen, VRM Chavali, GD Aguirre, WA Beltran
    Scientific Reports, 2021-01-11;11(1):460.
    Species: Human
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  4. Prolactin enhances interferon-gamma-induced production of CXC ligand 9 (CXCL9), CXCL10, and CXCL11 in human keratinocytes.
    Authors: Kanda N, Watanabe S
    Endocrinology, 2007-01-25;148(5):2317-25.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: ELISA Development
  5. Shift of monocyte function toward cellular immunity during sleep.
    Authors: Lange T, Dimitrov S, Fehm HL, Westermann J, Born J
    Arch. Intern. Med., 2006-09-18;166(16):1695-700.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  6. Prolactin and heregulin override DNA damage-induced growth arrest and promote phosphatidylinositol-3 kinase-dependent proliferation in breast cancer cells.
    Authors: Chakravarti P, Henry MK, Quelle FW
    Int. J. Oncol., 2005-02-01;26(2):509-14.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization

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