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    prame-antibody-epr20330-ab219650.pdf

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Tags & Cell Markers Cell Type Markers Tumor Associated
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RecombinantRabMAb

Recombinant Anti-PRAME antibody [EPR20330] (ab219650)

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  • Certificate of Compliance
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
  • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] (ab219650)
  • Flow Cytometry (Intracellular) - Anti-PRAME antibody [EPR20330] (ab219650)
  • Western blot - Anti-PRAME antibody [EPR20330] (ab219650)
  • Immunoprecipitation - Anti-PRAME antibody [EPR20330] (ab219650)
  • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] (ab219650)
  • Western blot - Anti-PRAME antibody [EPR20330] (ab219650)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
  • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] (ab219650)
  • Western blot - Anti-PRAME antibody [EPR20330] (ab219650)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
  • Anti-PRAME antibody [EPR20330] (ab219650)

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, IP, Flow Cyt (Intra)
  • Reacts with: Human

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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, IP, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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 (intra): 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

  • Form

    Liquid
  • Storage instructions

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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20330
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Cell Type Markers
    • Tumor Associated
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Co-activators/co-repressors
    • Cancer
    • Tumor biomarkers
    • Tumor antigens

Associated products

  • Alternative Versions

    • Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunohistochemistry kits

    • Rabbit specific HRP/DAB Detection IHC Detection Kit - Micro-polymer (ab236469)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human PRAME protein (ab132315)
  • Related Products

    • Anti-SOX10 antibody [EPR4007] (ab155279)
    • Anti-SOX10 antibody [EPR4007-104] (ab180862)
    • Anti-SOX10 antibody [SP267] (ab227680)
    • Anti-SOX10 antibody [SP275] (ab227684)

Applications

The Abpromise guarantee

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.
IP
1/30.
Flow Cyt (Intra)
1/500.
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.
IP
1/30.
Flow Cyt (Intra)
1/500.

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.
  • Target information above from: UniProt accession P78395 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 23532 Human
    • Omim: 606021 Human
    • SwissProt: P78395 Human
    • Unigene: 30743 Human
    • 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)

      ab219650 staining PRAME in K562 cells, with negative expression in Ramos cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab219650 at 1 µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 µg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

      This product also work with 100% methanol (5 min) fixation under the same testing conditions.

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

      Intracellular 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.

    • 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.

    • Western blot - Anti-PRAME antibody [EPR20330] (ab219650)
      Western blot - Anti-PRAME antibody [EPR20330] (ab219650)
      Different batches of ab219650 were tested on MeWo (Human malignant melanoma fibroblast) whole cell lysate at 0.1 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 50 kDa.
    • 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.

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

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (11)

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

    ab219650 has been referenced in 11 publications.

    • Donnell SA  et al. Comparison of adipophilin and recently introduced PReferentially expressed Antigen in MElanoma immunohistochemistry in the assessment of sebaceous neoplasms: A pilot study. J Cutan Pathol N/A:N/A (2021). PubMed: 33949693
    • Parra O  et al. Cross-reactivity of NRASQ61R antibody in a subset of Spitz nevi with 11p gain: a potential confounding factor in the era of pathway-based diagnostic approach. Hum Pathol 112:35-47 (2021). PubMed: 33636207
    • Zhang W  et al. Tumor-associated antigen Prame targets tumor suppressor p14/ARF for degradation as the  receptor protein of CRL2Prame complex. Cell Death Differ 28:1926-1940 (2021). PubMed: 33504946
    • Cadwell CR  et al. Preferentially Expressed Antigen in Melanoma (PRAME) Expression in Malignant, but Not Benign, Peripheral Nerve Sheath Tumors. J Neuropathol Exp Neurol 80:384-386 (2021). PubMed: 33212492
    • Liu Y  et al. Discovery of key genes as novel biomarkers specifically associated with HPV-negative cervical cancer. Mol Ther Methods Clin Dev 21:492-506 (2021). PubMed: 33997099
    View all Publications for this product

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