Overview

  • Product name

    Anti-Prolactin Receptor/PRL-R antibody [T6]
    See all Prolactin Receptor/PRL-R primary antibodies
  • Description

    Mouse monoclonal [T6] to Prolactin Receptor/PRL-R
  • Host species

    Mouse
  • Specificity

    Detects Prolactin Receptor/PRL-R in rat tissues. This antibody does not cross-react with Growth Hormone (GH) receptor. By Western blot, this antibody detects an ~42 kDa protein representing Prolactin Receptor/PRL-R in NB2 cell lysate. Immunohistochemical staining of Prolactin Receptor/PRL-R in NB2 cells yields a staining pattern consistent with cytoplasmic vesicular staining. This antibody has also been used to inhibit the binding of prolactin to Prolactin Receptor/PRL-R in vitro.

  • Tested applications

    Suitable for: IHC-P, ICC, IHC-Fr, WB, IP, Flow Cyt, Inhibition Assay, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Sheep, Human
  • Immunogen

    Other Immunogen Type corresponding to Rat Prolactin Receptor/PRL-R. Purified rat liver PRL receptor.

  • Positive control

    • WB and ICC: NB2 cells
  • General notes

    Previously labelled as Prolactin Receptor. 

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Primary antibody notes

    Prolactin (PRL) is a hormone involved in a variety of important functions including ion transport and osmoregulation, stimulation of milk, protein synthesis as well as the regulation of numerous reproductive functions. PRL exerts its influence on different cell types through a signal transduction pathway which begins with the binding of the hormone to a transmembrane PRL receptor. Immunoreactive PRL receptor, a member of the cytokine receptor family, varies in size (short and long forms) with tissue source and species, from ~40 kDa to 100 kDa. The PRL receptor consists of at least three separate domains: an extracellular region with 5 cysteines which contains the prolactin binding site, a single transmembrane domain and a cytoplasmic region, the length of which appears to influence ligand binding and regulate cellular function.
  • Clonality

    Monoclonal
  • Clone number

    T6
  • Isotype

    IgG2a
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2773 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/25 - 1/200.
ICC Use a concentration of 5 µg/ml.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

Inhibition Assay Use at an assay dependent concentration.
ICC/IF 1/200.

Target

  • Function

    This is a receptor for the anterior pituitary hormone prolactin (PRL). Isoform 4 is unable to transduce prolactin signaling. Isoform 6 is unable to transduce prolactin signaling.
  • Tissue specificity

    Expressed in breast, placenta, kidney, liver and pancreas.
  • Sequence similarities

    Belongs to the type I cytokine receptor family. Type 1 subfamily.
    Contains 2 fibronectin type-III domains.
  • Domain

    The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell-surface receptor binding.
    The box 1 motif is required for JAK interaction and/or activation.
  • Cellular localization

    Secreted and Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • AI987712 antibody
    • CLONE SPM213 antibody
    • CPRLP antibody
    • Delta 4-delta 7/11 truncated prolactin receptor antibody
    • Delta 4-SF1b truncated prolactin receptor antibody
    • HPRL antibody
    • hPRL receptor antibody
    • hPRLrI antibody
    • Lactogen receptor antibody
    • MFAB antibody
    • MGC105486 antibody
    • OPR antibody
    • OTTHUMP00000115998 antibody
    • Pr-1 antibody
    • Pr-3 antibody
    • PRL R antibody
    • PRL-R antibody
    • PRLR antibody
    • Prlr-rs1 antibody
    • PRLR_HUMAN antibody
    • Prolactin receptor a antibody
    • Prolactin receptor antibody
    • Prolactin receptor delta 7/11 antibody
    • RATPRLR antibody
    • Secreted prolactin binding protein antibody
    • Truncated testis-specific box 1-C prolactin receptor antibody
    • wu:fj65c07 antibody
    see all

Images

  • Immunocytochemistry/Immunofluorescence analysis of Prolactin Receptor/PRL-R in C6 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incbated without (control) or with ab2773 (1:200) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Prolactin Receptor/PRL-R staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Prolactin Receptor/PRL-R in H-4-II-E cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incbated without (control) or with ab2773 (1:200) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Prolactin Receptor/PRL-R staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Prolactin Receptor/PRL-R in SW480 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incbated without (control) or with ab2773 (1:200) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Prolactin Receptor/PRL-R staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) was performed on normal biopsies of deparaffinized rat pituitary gland tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and incubated with ab2773 (1:50) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

References

This product has been referenced in:

  • Dardenne M  et al. Prolactin receptor expression in human hematopoietic tissues analyzed by flow cytofluorometry. Endocrinology 134:2108-14 (1994). Read more (PubMed: 8156910) »
  • Rui H  et al. JAK2 activation and cell proliferation induced by antibody-mediated prolactin receptor dimerization. Endocrinology 135:1299-306 (1994). Read more (PubMed: 7925093) »
See all 6 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (liver, mammary gland)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer ph 6.0
Permeabilization
No
Specification
liver, mammary gland
Blocking step
protein block as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: rt°C
Fixative
Formaldehyde

Ms. Francine Deslippe

Verified customer

Submitted Apr 10 2013

Answer

Thank you for contacting us.
I am sorry to hear about the problems you have encountered.

The bands you are seeing at 25 kDa could be the light chain of endogenous IgG.

As we discussed over the phone, please try the following suggestions as it seems the WB conditions for these 2 antibodies (ab25630 and ab2773) are not yet optimal:

1) try blocking for only 1-2 h RT
2) try the primary antibody incubation for overnight 4 degree

As for ab25631, this has been shown to not react with rat samples and would probably not give satisfactory results.However, if you have access to a human sample, you could try the antibody on that.

Please let me know if the tips were helping to improve your results.

I hope this information is helpful to you, and I look forward to hear back.

Read More
Application
Western blot
Sample
Mouse Tissue lysate - whole (Forebrain)
Loading amount
40 µg
Specification
Forebrain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jan 09 2007

Question

Our customer has tried the proteolytic pretreatment with 0.1% trypsin but the result again was not very convincing (see attached pdf-file 'PrlR-Staining'). The file shows pictures taken from stainings with AB2773 (page2) and pictures taken from stainings with the non-commercial antibody T6 (page1, from S. Kloehn et al.). The T6 antibody is described in: Ouhtit A, Ronsin B, Kelly PA, Morel G. Ultrastructural expression of prolactin receptor in rat liver. Biol Cell. 1994;82(2-3):169-76 Okamura H, Zachwieja J, Raguet S, Kelly PA. Characterization and applications of monoclonal antibodies to the prolactin receptor. Endocrinology. 1989 May;124(5):2499-508. So you can see that the results obtained with AB2773 are still not very good. If you have any more suggestions on how the results can be imroved they will be highly appreciated. (Please notice that I am on holiday next week, but my collegue Christian Erlenhoefer will take my place during that time) Previous message: I have a complaint about your Prolactin Receptor antibody #AB2773: Our customer used it in IHC staining on formalin-fixed, paraffin embedded tissue sections of rat liver and pituitary. Protocol: Antigen Retrieval: - with 10 mM Citrate buffer pH 6.0 using a Microwave Blocking: 5 min 0.5% Triton in TBS wash with TBS 20-25 min 0-35% H2O2 in TBS/Methanol (90/10) wash 3x with TBS 10 min DAKO blocking solution A 10 min DAKO blocking solution B wash 3x with TBS Primary antibody: AB2773 (Lot 19068) 1/50 in antibody diluent (DAKO) for 1h at RT wash 3x with TBS Secondary antibody: biotinylated secondary antibody from DAKO (LSAB-Kit, ready to use) for 30 min at RT wash 3x with TBS Streptavidin-Biotin-Complex (DAKO) for 30 min at RT wash 3x with TBS Detection with AEC-Substrate Kit. Counterstaining in Hämalaun, embedding in Kaisers Glycine-Gelatine. Problem: no specific staining could be obtained. The same protocol was used successfully using a Ki67 antibody on rat tonsil. Please hava a look at the attached pictures.

Read More
Answer

Here is a reference that uses our monoclonal antibodies with IHC procedures: Rl Sorenson and LE Stout. Prolactin Receptors and JAK2 in Islets of Langerhans: An Immunohistochemical Analysis. Endocrinology 136:9 pp 4092-4098. 1995 Also used in FACS analysis: M. Gagnerault, et al. Expression of Prolactin Receptors in Murine Lymphoid Cells in Normal and Autoimmune Situations. J. Immunol., 150(12): 5673-5681, 1993. The customer should follow these protocols along with the other information have passed onto him/her with our previous correspondence. If not, please let us know how you would like to proceed. This will work, otherwise, there is another issue and not the antibody. Please let us know if this helps or if you require further assistance. Thank you,

Read More

Answer

Ab2773 has been shown to be specific for the short form around 42 kDa. The other Prolactin Receptor antibody that we currently have, ab2772, detects both the short and long forms. So unfortunately we don't have what your customer is looking for.

Read More

Answer

We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will forward a GBP10/ USD15/ EUR15 Amazon gift voucher.

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