Validated using a knockout cell line
Recombinant
RabMAb

Anti-FANCD2 antibody [EPR2302] (ab108928)

Overview

  • Product name
    Anti-FANCD2 antibody [EPR2302]
    See all FANCD2 primary antibodies
  • Description
    Rabbit monoclonal [EPR2302] to FANCD2
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues in Human FANCD2.

  • Positive control
    • WB: HAP1, HeLa, MCF7, SKBR-3, K562, HL-60, C6, and PC-12 cell lysates. IHC-P: Human tonsil tissue. ICC/IF: HAP1 and HeLa cells. Flow Cyt: Jurkat cells.
  • General notes

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

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. Stable for 12 months at -20°C.
  • Storage buffer
    PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Purity
    Tissue culture supernatant
  • Clonality
    Monoclonal
  • Clone number
    EPR2302
  • Isotype
    IgG
  • Research areas

Applications

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
Flow Cyt 1/2000.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/250.
IP 1/10 - 1/100.
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.
  • Information by UniProt
  • Database links
  • 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

Anti-FANCD2 antibody [EPR2302] images



  • Predicted band size : 166 kDa

    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 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.

  • ab108928 staining FANCD2 in wild-type HAP1 cells (top panel) and FANCD2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), 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 1h. 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 1h 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.

  • Immunohistochemical staining of paraffin-embedded Human tonsil tissue using ab108928 at a dilution of 1/100.
  • Flow cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling FANCD2 (red) with ab108928 at a 1/2000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 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.

  • ab108928 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

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

    Lane 1 : HeLa cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : SKBR-3 cell lysate
    Lane 4 : K562 cell lysate
    Lane 5 : HL60 cell lysate
    Lane 6 : C6 cell lysate
    Lane 7 : PC-12 cell lysate

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 166 kDa
  • Immunofluorescent staining of HeLa cells using ab108928 at a dilution of 1/250.

References for Anti-FANCD2 antibody [EPR2302] (ab108928)

This product has been referenced in:
  • Serber DW  et al. The Mouse INO80 Chromatin-Remodeling Complex Is an Essential Meiotic Factor for Spermatogenesis. Biol Reprod 94:8 (2016). ICC/IF ; Mouse . Read more (PubMed: 26607718) »
  • Jiang Q  et al. MERIT40 cooperates with BRCA2 to resolve DNA interstrand cross-links. Genes Dev 29:1955-68 (2015). Human . Read more (PubMed: 26338419) »

See all 9 Publications for this product

Product Wall

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
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Verified customer

Submitted Apr 25 2014

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (6%)
Sample
Mouse Cell lysate - whole cell (Mouse Pancreatic cancer)
Specification
Mouse Pancreatic cancer
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Mar 18 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (6%)
Sample
Human Tissue lysate - whole (U2OS osteosarcoma)
Specification
U2OS osteosarcoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Mar 04 2014

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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