Recombinant
RabMAb

Recombinant Anti-PRAME antibody [EPR20330] (ab219650)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20330] to PRAME
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-PRAME antibody [EPR20330]
    See all PRAME primary antibodies

  • Description

    Rabbit monoclonal [EPR20330] to PRAME

  • Host species

    Rabbit

  • Specificity

    PRAME is expressed in malignant cells, including leukaemias, Hodgkin's lymphoma, breast cancer, and primary and metastatic melanomas.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human PRAME aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: P78395

  • Positive control

    • WB: MeWo and A-375 whole cell lysates; Human ovary cancer and testis lysates. IHC-P: Human testis and melanoma tissues. ICC/IF: MeWo and A-375 cells. Flow Cyt: MeWo cells. IP: MeWo whole cell lysate.

  • General notes

    PRAME (PReferentially expressed Antigen in MElanoma) is a tumor-associated antigen and is a member of the family of cancer testis antigens (CTA). PRAME is expressed in malignant cells, including leukaemias, Hodgkin's lymphoma, breast cancer, and primary and metastatic melanomas. For more information, please refer to PMID: 27441500. PRAME has low or no expression in normal tissues except for in testis, ovary, placenta, adrenals and endometrium. For more information, please refer to PMID: 9047241.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab219650 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
WB 1/1000. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa).
IHC-P 1/16000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
Flow Cyt 1/500.
IP 1/30.

Target

  • Function

    Functions as a transcriptional repressor, inhibiting the signaling of retinoic acid through the retinoic acid receptors RARA, RARB and RARG. Prevents retinoic acid-induced cell proliferation arrest, differentiation and apoptosis.

  • Tissue specificity

    Expressed in testis. Detected in samples of kidney, brain and skin.

  • Sequence similarities

    Belongs to the PRAME family.
    Contains 4 LRR (leucine-rich) repeats.

  • Cellular localization

    Nucleus. Cell membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • 4930534P07Rik antibody
      • Cancer/testis antigen 130 antibody
      • CT130 antibody
      • MAPE antibody
      • Melanoma antigen preferentially expressed in tumors antibody
      • OIP 4 antibody
      • OIP-4 antibody
      • OIP4 antibody
      • OPA interacting protein 4 antibody
      • Opa interacting protein OIP4 antibody
      • OPA-interacting protein 4 antibody
      • PRAME antibody
      • PRAME_HUMAN antibody
      • Preferentially expressed antigen in melanoma antibody
      • Preferentially expressed antigen of melanoma antibody
      • RP23-250F8.3 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)

      Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling PRAME with ab219650 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on human melanoma (PMID: 9047241).

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] (ab219650)
      Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] (ab219650)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MeWo (Human malignant melanoma cell line) cells labeling PRAME with ab219650 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mostly nuclear staining on MeWo cells.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    • Flow Cytometry - Anti-PRAME antibody [EPR20330] (ab219650)
      Flow Cytometry - Anti-PRAME antibody [EPR20330] (ab219650)

      Flow cytometric analysis of 4% paraformaldehyde-fixed MeWo (Human malignant melanoma cell line) cells labeling PRAME with ab219650 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    • Western blot - Anti-PRAME antibody [EPR20330] (ab219650)
      Western blot - Anti-PRAME antibody [EPR20330] (ab219650)
      All lanes : Anti-PRAME antibody [EPR20330] (ab219650) at 1/1000 dilution

      Lane 1 : Human ovary cancer lysate
      Lane 2 : A-375 (Human malignant melanoma cell line) whole cell lysate
      Lane 3 : Human testis lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Predicted band size: 57 kDa
      Observed band size: 57 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1: 3 minutes; Lane 2: 5 seconds; Lane 3: 1 minute.

    • Immunoprecipitation - Anti-PRAME antibody [EPR20330] (ab219650)
      Immunoprecipitation - Anti-PRAME antibody [EPR20330] (ab219650)

      PRAME was immunoprecipitated from 0.35 mg of MeWo (Human malignant melanoma cell line) whole cell lysate with ab219650 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219650 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: MeWo whole cell lysate 10 μg (Input).

      Lane 2: ab219650 IP in MeWo whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219650 in MeWo whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 1 second.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)

      Immunohistochemical analysis of paraffin-embedded human testis tissue labeling PRAME with ab219650 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on human testis (PMID: 9047241).

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] (ab219650)
      Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] (ab219650)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A-375 (Human malignant melanoma cell line) cells labeling PRAME with ab219650 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mostly nuclear staining on A-375 cells.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    • Western blot - Anti-PRAME antibody [EPR20330] (ab219650)
      Western blot - Anti-PRAME antibody [EPR20330] (ab219650)
      Anti-PRAME antibody [EPR20330] (ab219650) at 1/1000 dilution + MeWo (Human malignant melanoma cell line) whole cell lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/4000 dilution

      Predicted band size: 57 kDa
      Observed band size: 57 kDa


      Exposure time: 15 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)

      Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling PRAME with ab219650 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Negative control: No staining on human pancreas (PMID: 9047241).

      Counter stained with Hematoxylin.

       

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    References

    Publishing research using ab219650? Please let us know so that we can cite the reference in this datasheet.

    ab219650 has been referenced in 4 publications.

    • Lezcano C  et al. Immunohistochemistry for PRAME in the Distinction of Nodal Nevi From Metastatic Melanoma. Am J Surg Pathol 44:503-508 (2020). PubMed: 31633488
    • Hodgson A  et al. Evaluation of cancer testis antigen (CT10, PRAME) and MHC I expression in high-grade urothelial carcinoma of the bladder. Virchows Arch 476:535-542 (2020). PubMed: 31485721
    • Toyama A  et al. Analyses of molecular and histopathologic features and expression of PRAME by immunohistochemistry in mucosal melanomas. Mod Pathol N/A:N/A (2019). PubMed: 31375769
    • Lezcano C  et al. PRAME Expression in Melanocytic Tumors. Am J Surg Pathol 42:1456-1465 (2018). PubMed: 30045064

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