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RecombinantRabMAb

Recombinant Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
  • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
  • Western blot - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
  • Flow Cytometry (Intracellular) - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
  • Immunoprecipitation - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
  • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
  • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
  • Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)

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 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-P
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Unconjugated

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HRP Conjugation Kit - Lightning-Link® (ab102890)
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Overview

  • Product name

    Anti-PRAME antibody [EPR20330] - BSA and Azide free
    See all PRAME primary antibodies
  • Description

    Rabbit monoclonal [EPR20330] to PRAME - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • IHC-P: Human melanoma tissue.
  • General notes

    ab232571 is the carrier-free version of ab219650.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • 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] (ab219650)
    • Alexa Fluor® 555 Anti-PRAME antibody [EPR20330] (ab307626)
    • Alexa Fluor® 647 Anti-PRAME antibody [EPR20330] (ab307630)
    • Alexa Fluor® 488 Anti-PRAME antibody [EPR20330] (ab307769)
  • Compatible Secondaries

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

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

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

    • Recombinant Human PRAME protein (ab132315)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Anti-SOX10 antibody [EPR4007] - BSA and Azide free (ab186821)
    • Anti-SOX10 antibody [EPR4007-104] - BSA and Azide free (ab220078)
    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab232571 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa).
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Recommend ab219650 incubation at +4°C overnight.

Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa).
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Recommend ab219650 incubation at +4°C overnight.

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] - BSA and Azide free (ab232571)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)

      Immunohistochemical analysis of paraffin-embedded sections of Human melanoma tissue labelling PRAME with ab219650 at 1/500 dilution (1 μg/ml) followed by a Goat Anti-Rabbit IgG H&L (HRP polymer) secondary antibody at a ready to use concentration.

      Positive staining on human melanoma tissue.

      The section was incubated with ab219650 at +4°C overnight. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)

      Immunohistochemical analysis of paraffin-embedded sections of Human tonsil tissue labelling PRAME with at 1/500 dilution (1 μg/ml) followed by a Goat Anti-Rabbit IgG H&L (HRP polymer) secondary antibody at a ready to use concentration.

      Negative control: no staining on human tonsil.

      The section was incubated with ab219650 at +4°C overnight. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).

    • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
      Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    • Western blot - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
      Western blot - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
      This data was developed using ab219650, the same antibody clone in a different buffer formulation. 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.
    • Flow Cytometry (Intracellular) - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
      Flow Cytometry (Intracellular) - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed and 90% Methanol-permeabilised 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. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650). 

       

       

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

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).

    • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
      Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).

    • Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)
      Immunocytochemistry/ Immunofluorescence - Anti-PRAME antibody [EPR20330] - BSA and Azide free (ab232571)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219650).

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

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

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

    References (0)

    Publishing research using ab232571? Please let us know so that we can cite the reference in this datasheet.

    ab232571 has not yet been referenced specifically in any publications.

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