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  1. Link

    folate-binding-proteinfbp-antibody-epr20277-ab221543.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
  • Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
  • Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
  • Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
  • Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20277] to Folate Binding Protein/FBP
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Secondary
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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Knockout
Product image
Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (ab264921)

View more associated products

Overview

  • Product name

    Anti-Folate Binding Protein/FBP antibody [EPR20277]
    See all Folate Binding Protein/FBP primary antibodies
  • Description

    Rabbit monoclonal [EPR20277] to Folate Binding Protein/FBP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal kidney, mouse kidney, and rat kidney tissue lysates and JAR, and HeLa cell lysates. IHC-P: Human kidney, ovarian cancer and lung cancer tissues; mouse kidney tissue; rat kidney tissue.
  • General notes

    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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20277
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Cell Type Markers
    • Tumor Associated

Associated products

  • Alternative Versions

    • Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free (ab235140)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (ab264921)
  • KO cell lysates

    • Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell lysate (ab257270)
  • Related Products

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab221543 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 - 1/5000. Detects a band of approximately 38 kDa (predicted molecular weight: 30 kDa).
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
WB
1/1000 - 1/5000. Detects a band of approximately 38 kDa (predicted molecular weight: 30 kDa).
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Binds to folate and reduced folic acid derivatives and mediates delivery of 5-methyltetrahydrofolate to the interior of cells.
  • Tissue specificity

    Exclusively expressed in tissues of epithelial origin. Expression is increased in malignant tissues. Expressed in kidney, lung and cerebellum.
  • Involvement in disease

    Defects in FOLR1 are the cause of neurodegeneration due to cerebral folate transport deficiency (NCFTD) [MIM:613068]. NCFTD is an autosomal recessive disorder resulting from brain-specific folate deficiency early in life. Onset is apparent in late infancy with severe developmental regression, movement disturbances, epilepsy, and leukodystrophy. Note=Recognition and diagnosis of this disorder is critical because folinic acid therapy can reverse the clinical symptoms and improve brain abnormalities and function.
  • Sequence similarities

    Belongs to the folate receptor family.
  • Post-translational
    modifications

    Eight disulfide bonds are present.
    The secreted form is derived from the membrane-bound form either by cleavage of the GPI anchor, or/and by proteolysis catalyzed by a metalloprotease.
  • Cellular localization

    Cell membrane. Secreted.
  • Target information above from: UniProt accession P15328 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2348 Human
    • Entrez Gene: 171049 Rat
    • Omim: 136430 Human
    • SwissProt: P15328 Human
    • SwissProt: P35846 Mouse
    • Unigene: 73769 Human
    • Unigene: 2135 Mouse
    • Alternative names

      • adult antibody
      • Adult folate binding protein antibody
      • Adult folate-binding protein antibody
      • FBP antibody
      • Folate Binding Protein antibody
      • Folate Receptor 1 Adult antibody
      • Folate receptor 1 antibody
      • Folate Receptor 1 Precursor antibody
      • Folate receptor adult antibody
      • Folate receptor alpha antibody
      • Folate receptor antibody
      • FOLR antibody
      • FOLR1 antibody
      • FOLR1_HUMAN antibody
      • FR alpha antibody
      • FR-alpha antibody
      • FRalpha antibody
      • KB cells FBP antibody
      • MOV18 antibody
      • Ovarian cancer associated antigen antibody
      • Ovarian tumor associated antigen antibody
      • Ovarian tumor associated antigen MOv18 antibody
      • Ovarian tumor-associated antigen MOv18 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)

      Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051, ready to use). Cytoplasmic staining on human kidney (PMID: 22204844). Counter stained with Hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051, ready to use).

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

    • Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      All lanes : Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : FOLR1 knockout HeLa cell lysate
      Lane 3 : JAR cell lysate
      Lane 4 : MDA-MB-231 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 30 kDa
      Observed band size: 38 kDa why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab221543 observed at 38 kDa. Red - loading control, ab7291 observed at 52 kDa.

      ab221543 Anti-Folate Binding Protein/FBP antibody [EPR20277] was shown to specifically react with Folate Binding Protein/FBP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264921 (knockout cell lysate ab257270) was used. Wild-type and Folate Binding Protein/FBP knockout samples were subjected to SDS-PAGE. ab221543 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

       

    • Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      All lanes : Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543) at 1/5000 dilution

      Lane 1 : GST-His-tagged human recombinant FOLR1 (aa 25-160), 20 ng
      Lane 2 : His-tagged human recombinant FOLR2 (aa 24-154), 20 ng

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 30 kDa
      Observed band size: 40 kDa why is the actual band size different from the predicted?



      Exposure time:

      Lanes 1-2: 5 seconds

    • Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      All lanes : Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543) at 1/1000 dilution

      Lane 1 : JAR (human placenta choriocarcinoma epithelial cells) whole cell lysate at 20 µg
      Lane 2 : Human fetal kidney lysate at 20 µg
      Lane 3 : Mouse kidney lysate at 20 µg
      Lane 4 : Rat kidney lysate
      Lane 5 : HeLa (Human cervix adenocarcinoma epithelial cells) whole cell lysate at 10 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 30 kDa
      Observed band size: 38 kDa why is the actual band size different from the predicted?



      Exposure times:

      Lane 1:10 seconds

      Lane 2: 30 seconds

      Lanes 3-4: 5 seconds

      Lane 5: 3 minutes

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)

      Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). Mainly membranous staining on tumor cells of human ovarian cancer (PMID: 22204844). Counter stained with Hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use).

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)

      Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). Cytoplasmic staining on tumor cells of human lung cancer (PMID: 22204844). Counter stained with Hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use).

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)

      Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). Positive staining on mouse kidney (PMID: 22204844). Counter stained with Hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use).

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)

      Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). Positive staining on rat kidney (PMID: 22204844). Counter stained with Hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use).

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)

      Negative control:

      Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). No staining on human tonsil (PMID: 22547449). Counter stained with Hematoxylin.

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

    • Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)
      Anti-Folate Binding Protein/FBP antibody [EPR20277] (ab221543)

    Protocols

    • Western blot protocols
    • Immunohistochemistry 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 (4)

    Publishing research using ab221543? Please let us know so that we can cite the reference in this datasheet.

    ab221543 has been referenced in 4 publications.

    • Furugen A  et al. Effects of valproate, an HDAC inhibitor, on the expression of folate carriers and folate metabolism-related genes in the placenta of rats. Drug Metab Pharmacokinet 40:100409 (2021). PubMed: 34293696
    • Wang Y  et al. Decitabine Sensitizes the Radioresistant Lung Adenocarcinoma to Pemetrexed Through Upregulation of Folate Receptor Alpha. Front Oncol 11:668798 (2021). PubMed: 34079760
    • Maniati E  et al. Mouse Ovarian Cancer Models Recapitulate the Human Tumor Microenvironment and Patient Response to Treatment. Cell Rep 30:525-540.e7 (2020). PubMed: 31940494
    • Jin YH  et al. Diagnostic Value of ER, PR, FR and HER-2-Targeted Molecular Probes for Magnetic Resonance Imaging in Patients with Breast Cancer. Cell Physiol Biochem 49:271-281 (2018). PubMed: 30138940

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