Human Prolactin DuoSet ELISA DY682: R&D Systems

Human Prolactin DuoSet ELISA

(6 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Sample Type & Volume Required
    100 µL
  • Range
    15.60 - 1,000 pg/mL
  • Sufficient Materials
    For fifteen 96-well plates*
  • Specificity
    Please see the product datasheet
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
Ancillary Reagent Kit Available

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Prolactin. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.


Product Features
  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits
Kit Content
  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Prolactin
Prolactin (PRL) is a neuroendocrine pituitary hormone that is synthesized by anterior pituitary, placenta, brain, uterus, dermal fibroblasts, decidua, B cells, T cells, natural killer cells, and breast cancer cells. The prolactin receptor is a transmembrane type I glycoprotein that belongs to the hematopoietic cytokine receptor family.

Prolactin receptor (PRL R) contains an extracellular, transmembrane, and intracellular domain. Transcriptional regulation of the PRL R gene results in several different species-dependent isoforms of PRL R being produced.

  • Entrez Gene IDs:
    5617 (Human); 19109 (Mouse);
  • Aliases:
    PRL; Prolactin
Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations:

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
Filter your results:

Species
Sample Type
  1. Decidualization is Impaired in Endometrial Stromal Cells from Uterine Rudiments in Mayer-Rokitansky-K�ster-Hauser Syndrome
    Authors: SY Brucker, S Eisenbeis, J König, M Lamy, MS Salker, N Zeng, H Seeger, M Henes, D Schöller, B Schönfisch, A Staebler, FA Taran, D Wallwiener, K Rall
    Cell. Physiol. Biochem, 2017;41(3):1083-1097.
    Species: Human
    Sample Type: Cell Culture Supernates
  2. Cardiac angiogenic imbalance leads to peripartum cardiomyopathy.
    Authors: Patten IS, Rana S, Shahul S, Rowe GC, Jang C, Liu L, Hacker MR, Rhee JS, Mitchell J, Mahmood F, Hess P, Farrell C, Koulisis N, Khankin EV, Burke SD, Tudorache I, Bauersachs J, del Monte F, Hilfiker-Kleiner D, Karumanchi SA, Arany Z
    Nature, 2012;485(7398):333-8.
    Species: Human
    Sample Type: Plasma
  3. Calcitriol stimulates prolactin expression in non-activated human peripheral blood mononuclear cells: breaking paradigms.
    Authors: Diaz L, Martinez-Reza I, Garcia-Becerra R, Gonzalez L, Larrea F, Mendez I
    Cytokine, 2011;55(2):188-94.
    Species: Human
    Sample Type: Cell Culture Supernates
  4. Heparin and low-molecular-weight heparins modulate the decidualization of human endometrial stromal cells.
    Authors: Fluhr H, Spratte J, Ehrhardt J, Steinmuller F, Licht P, Zygmunt M
    Fertil. Steril., 2010;93(8):2581-7.
    Species: Human
    Sample Type: Cell Culture Supernates
  5. Manipulating actin dynamics affects human in vitro decidualization.
    Authors: Ihnatovych I, Livak M, Reed J, de Lanerolle P, Strakova Z
    Biol. Reprod., 2009;81(1):222-30.
    Species: Human
    Sample Type: Cell Culture Supernates
  6. Reduction in hypophyseal growth hormone and prolactin expression due to deficiency in ghrelin receptor signaling is associated with Pit-1 suppression: Relevance to the immune system.
    Authors: Yang H, Dixit VD, Patel K, Vandanmagsar B, Collins G, Sun Y, Smith RG, Taub DD
    Brain Behav. Immun., 2008;0(0):.
    Species: Mouse
    Sample Type: Serum
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DuoSet ELISA Ancillary Reagent Kit 2

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Stop Solution 2N Sulfuric Acid (15 x 6 mL)

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ELISA Plate Sealers, 100 Pack

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Streptavidin-HRP (set of 5 vials enough for 75 plates total)

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Quantikine Wash Buffer 1

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Clear Polystyrene Microplates, 25 Pack

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Glo Substrate Reagent Pack (Reagent A & B)

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ELISA Plate-coating Buffer (250 mL of PBS)

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Reagent Diluent Concentrate 3 (5X, 10 x 21 mL)

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Black Polystyrene Microplates, 25 Pack

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