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  1. Link

    fancd2-antibody-epr2302-ab108928.pdf

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Epigenetics and Nuclear Signaling DNA / RNA DNA Damage & Repair Fanconi's Anemia
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-FANCD2 antibody [EPR2302] (ab108928)

  • Datasheet
  • SDS
Reviews (4) Submit a question References (32)

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Western blot - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Western blot - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Western blot - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Immunoprecipitation - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Flow Cytometry (Intracellular) - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)
  • Anti-FANCD2 antibody [EPR2302] (ab108928)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2302] to FANCD2
  • Suitable for: IHC-P, ICC/IF, IP, Flow Cyt (Intra), WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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Alexa Fluor® 488 Anti-FANCD2 antibody [EPR2302] (ab200759)
Knockout
Product image
Human FANCD2 knockout HeLa cell line (ab261743)

View more associated products

Overview

  • Product name

    Anti-FANCD2 antibody [EPR2302]
    See all FANCD2 primary antibodies
  • Description

    Rabbit monoclonal [EPR2302] to FANCD2
  • Host species

    Rabbit
  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • WB: HAP1, HeLa and HEK293 cell lysates. IHC-P: Human tonsil tissue. ICC/IF: HAP1 and HeLa cells. Flow Cyt (intra): Jurkat cells. IP: HeLa cell lysate
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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: 7.20
    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

    EPR2302
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • Fanconi's Anemia

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-FANCD2 antibody [EPR2302] (ab200759)
    • Alexa Fluor® 647 Anti-FANCD2 antibody [EPR2302] (ab200763)
    • Anti-FANCD2 antibody [EPR2302] - BSA and Azide free (ab221932)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

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

    • Human FANCD2 knockout HeLa cell line (ab261743)
  • KO cell lysates

    • Human FANCD2 knockout HeLa cell lysate (ab257173)
  • KO cell pellets

    • Human FANCD2 knockout HeLa cell pellet (ab278841)
  • Positive Controls

    • PC-12 nuclear extract lysate (ab14884)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108928 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
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

 

 

See IHC antigen retrieval protocols.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

ICC/IF (1)
1/250 - 1/500.
IP
1/10 - 1/100.
Flow Cyt (Intra)
1/200.
WB (3)
1/1000 - 1/10000. Detects a band of approximately 155 kDa (predicted molecular weight: 166 kDa).
Notes
IHC-P
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

 

 

See IHC antigen retrieval protocols.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

ICC/IF
1/250 - 1/500.
IP
1/10 - 1/100.
Flow Cyt (Intra)
1/200.
WB
1/1000 - 1/10000. Detects a band of approximately 155 kDa (predicted molecular weight: 166 kDa).

Target

  • Function

    Required for maintenance of chromosomal stability. Promotes accurate and efficient pairing of homologs during meiosis. Involved in the repair of DNA double-strand breaks, both by homologous recombination and single-strand annealing. May participate in S phase and G2 phase checkpoint activation upon DNA damage. Promotes BRCA2/FANCD1 loading onto damaged chromatin. May also be involved in B-cell immunoglobulin isotype switching.
  • Tissue specificity

    Highly expressed in germinal center cells of the spleen, tonsil, and reactive lymph nodes, and in the proliferating basal layer of squamous epithelium of tonsil, esophagus, oropharynx, larynx and cervix. Expressed in cytotrophoblastic cells of the placenta and exocrine cells of the pancreas (at protein level). Highly expressed in testis, where expression is restricted to maturing spermatocytes.
  • Involvement in disease

    Defects in FANCD2 are a cause of Fanconi anemia complementation group D type 2 (FANCD2) [MIM:227646]. It is a disorder affecting all bone marrow elements and resulting in anemia, leukopenia and thrombopenia. It is associated with cardiac, renal and limb malformations, dermal pigmentary changes, and a predisposition to the development of malignancies. At the cellular level it is associated with hypersensitivity to DNA-damaging agents, chromosomal instability (increased chromosome breakage) and defective DNA repair.
  • Developmental stage

    Highly expressed in fetal oocytes, and in hematopoietic cells of the fetal liver and bone marrow (at protein level).
  • Domain

    The C-terminal 24 residues of isoform 2 are required for its function.
  • Post-translational
    modifications

    Monoubiquitinated on Lys-561 during S phase and upon genotoxic stress (isoform 1 and isoform 2). Deubiquitinated by USP1 as cells enter G2/M, or once DNA repair is completed. Monoubiquitination requires the FANCA-FANCB-FANCC-FANCE-FANCF-FANCG-FANCM complex, RPA1 and ATR, and is mediated by FANCL/PHF9. Ubiquitination is required for binding to chromatin, interaction with BRCA1, BRCA2 and MTMR15/FAN1, DNA repair, and normal cell cycle progression, but not for phosphorylation on Ser-222 or interaction with MEN1.
    Phosphorylated in response to various genotoxic stresses by ATM and/or ATR. Upon ionizing radiation, phosphorylated by ATM on Ser-222 and Ser-1404. Phosphorylation on Ser-222 is required for S-phase checkpoint activation, but not for ubiquitination, foci formation, or DNA repair. In contrast, phosphorylation by ATR on other sites may be required for ubiquitination and foci formation.
  • Cellular localization

    Nucleus. Concentrates in nuclear foci during S phase and upon genotoxic stress.
  • Target information above from: UniProt accession Q9BXW9 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2177 Human
    • Entrez Gene: 211651 Mouse
    • Entrez Gene: 312641 Rat
    • Omim: 613984 Human
    • SwissProt: Q9BXW9 Human
    • SwissProt: Q80V62 Mouse
    • SwissProt: Q6IV68 Rat
    • Unigene: 208388 Human
    • Alternative names

      • DKFZp762A223 antibody
      • FA 4 antibody
      • FA D2 antibody
      • FA4 antibody
      • FAC D2 antibody
      • FACD 2 antibody
      • FACD antibody
      • FACD2 antibody
      • FACD2_HUMAN antibody
      • FAD antibody
      • FAD2 antibody
      • FANC D2 antibody
      • FANCD 2 antibody
      • FANCD antibody
      • FANCD2 antibody
      • FANCONI ANEMIA COMPLEMENTATION GROUP D antibody
      • Fanconi anemia complementation group D2 antibody
      • Fanconi anemia group D2 protein antibody
      • FANCONI PANCYTOPENIA TYPE 4 antibody
      • FLJ23826 antibody
      • OTTHUMP00000158853 antibody
      • OTTHUMP00000207925 antibody
      • Protein FACD2 antibody
      • Type 4 Fanconi pancytopenia antibody
      see all

    Images

    • Western blot - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Western blot - Anti-FANCD2 antibody [EPR2302] (ab108928)
      All lanes : Anti-FANCD2 antibody [EPR2302] (ab108928) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : FANCD2 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 166 kDa
      Observed band size: 166 kDa



      Lanes 1- 2: Merged signal (red and green). Green - ab108928 observed at 166 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

       ab108928 was shown to react with FANCD2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261743 (knockout cell lysate ab257173) was used. Wild-type HeLa and FANCD2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108928 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)

      ab108928 (unpurified) staining FANCD2 in wild-type HAP1 cells (top panel) and FANCD2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10 minutes), permeabilized 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 1 hour. The cells were then incubated with ab108928 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling FANCD2 with purified ab108928 at 1/50 dilution (3.6 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Western blot - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Western blot - Anti-FANCD2 antibody [EPR2302] (ab108928)
      All lanes : Anti-FANCD2 antibody [EPR2302] (ab108928) at 1/1000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
      Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysates
      Lane 4 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
      Lane 5 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 166 kDa
      Observed band size: 165 kDa why is the actual band size different from the predicted?

    • Western blot - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Western blot - Anti-FANCD2 antibody [EPR2302] (ab108928)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: FANCD2 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: HEK293 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab108928 (unpurified) observed at 165 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab108928 was shown to specifically react with FANCD2 when FANCD2 knockout samples were used. Wild-type and FANCD2 knockout samples were subjected to SDS-PAGE. ab108928 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)

      Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling FANCD2 with purified ab108928 at 1/500 dilution (0.4 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Immunoprecipitation - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Immunoprecipitation - Anti-FANCD2 antibody [EPR2302] (ab108928)

      ab108928 (purified) at 1/20 dilution (1ug) immunoprecipitating FANCD2 in HeLa whole cell lysates.
      Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates 10ug
      Lane 2 (+): ab108928 & HeLa whole cell lysates
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108928 in HeLa whole cell lysates
      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Flow Cytometry (Intracellular) - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Flow Cytometry (Intracellular) - Anti-FANCD2 antibody [EPR2302] (ab108928)

      Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling FANCD2 (red) with ab108928 (purified) at a 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with primary and secondary antibodies. 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCD2 antibody [EPR2302] (ab108928)

      Immunohistochemical staining of paraffin-embedded human tonsil tissue using ab108928 (unpurified) at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)This image is courtesy of an anonymous Abreview

      ab108928 (unpurified) staining FANCD2 in human U2OS osteosarcoma cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 10% goat serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/300) for 18 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG monoclonal (1/1000) was used as the secondary antibody.

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)
      Immunocytochemistry/ Immunofluorescence - Anti-FANCD2 antibody [EPR2302] (ab108928)

      HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for FANCD2 (green) using ab108928 (unpurified) (1/250 dilution) in ICC/IF.

    • Anti-FANCD2 antibody [EPR2302] (ab108928)
      Anti-FANCD2 antibody [EPR2302] (ab108928)

    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

    References (32)

    Publishing research using ab108928? Please let us know so that we can cite the reference in this datasheet.

    ab108928 has been referenced in 32 publications.

    • Kaur A  et al. WNT inhibition creates a BRCA-like state in Wnt-addicted cancer. EMBO Mol Med 13:e13349 (2021). PubMed: 33660437
    • Gundogdu R  et al. hMOB2 deficiency impairs homologous recombination-mediated DNA repair and sensitises cancer cells to PARP inhibitors. Cell Signal 87:110106 (2021). PubMed: 34363951
    • Tan W  et al. Monoubiquitination by the human Fanconi anemia core complex clamps FANCI:FANCD2 on DNA in filamentous arrays. Elife 9:N/A (2020). PubMed: 32167469
    • Kreutmair S  et al. Loss of the Fanconi anemia-associated protein NIPA causes bone marrow failure. J Clin Invest 130:2827-2844 (2020). PubMed: 32338640
    • Sharp MF  et al. Methodology for the identification of small molecule inhibitors of the Fanconi Anaemia ubiquitin E3 ligase complex. Sci Rep 10:7959 (2020). PubMed: 32409752
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunocytochemistry/ Immunofluorescence abreview for Anti-FANCD2 antibody [EPR2302]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Sample
    Human Cell (u2os osteosarcoma)
    Specification
    u2os osteosarcoma
    Permeabilization
    Yes - 0.5% triton
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 25 2014

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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