重组Anti-FANCD2抗体[EPR2302] - BSA and Azide free (ab221932)
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 - BSA and Azide free
- Suitable for: IP, ICC/IF, WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-FANCD2抗体[EPR2302] - BSA and Azide free
参阅全部 FANCD2 一抗 -
描述
兔单克隆抗体[EPR2302] to FANCD2 - BSA and Azide free -
宿主
Rabbit -
特异性
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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经测试应用
适用于: IP, ICC/IF, WB, IHC-P, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, Hap1, MCF7, SKBR-3, K562, HL-60, C6, HEK293 and PC-12 cell lysates. IHC-P: Human tonsil tissue. ICC/IF: HeLa and wild-type HAP1 cells. IP: HeLa cell lysate
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常规说明
ab221932 is the carrier-free version of ab108928.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR2302 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-FANCD2 antibody [EPR2302] (ab108928)
- Alexa Fluor® 488 Anti-FANCD2 antibody [EPR2302] (ab200759)
- Alexa Fluor® 647 Anti-FANCD2 antibody [EPR2302] (ab200763)
- PE Anti-FANCD2 antibody [EPR2302] (ab307174)
- APC Anti-FANCD2 antibody [EPR2302] (ab307175)
- HRP Anti-FANCD2 antibody [EPR2302] (ab307176)
- Alexa Fluor® 594 Anti-FANCD2 antibody [EPR2302] (ab310563)
- Alexa Fluor® 555 Anti-FANCD2 antibody [EPR2302] (ab312092)
- Alexa Fluor® 568 Anti-FANCD2 antibody [EPR2302] (ab312572)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab221932于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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IP |
Use at an assay dependent concentration.
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ICC/IF | (1) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 155 kDa (predicted molecular weight: 166 kDa).
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IHC-P |
Use at an assay dependent concentration. 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. |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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说明 |
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IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 155 kDa (predicted molecular weight: 166 kDa). |
IHC-P
Use at an assay dependent concentration. 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. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
靶标
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功能
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. -
组织特异性
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. -
疾病相关
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. -
发展阶段
Highly expressed in fetal oocytes, and in hematopoietic cells of the fetal liver and bone marrow (at protein level). -
结构域
The C-terminal 24 residues of isoform 2 are required for its function. -
翻译后修饰
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. -
细胞定位
Nucleus. Concentrates in nuclear foci during S phase and upon genotoxic stress. - Information by UniProt
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数据库链接
- 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
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别名
- DKFZp762A223 antibody
- FA 4 antibody
- FA D2 antibody
see all
图片
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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 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab108928).
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.
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108928).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108928).
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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.
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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 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108928).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108928).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108928).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108928).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108928).
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Immunohistochemical staining of paraffin-embedded human tonsil tissue using ab108928 (unpurified) at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108928).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (10)
ab221932 被引用在 10 文献中.
- Siegner SM et al. Adenine base editing efficiently restores the function of Fanconi anemia hematopoietic stem and progenitor cells. Nat Commun 13:6900 (2022). PubMed: 36371486
- Richardson CD et al. CRISPR-Cas9 genome editing in human cells occurs via the Fanconi anemia pathway. Nat Genet 50:1132-1139 (2018). PubMed: 30054595
- Ramirez YP et al. Evaluation of novel imidazotetrazine analogues designed to overcome temozolomide resistance and glioblastoma regrowth. Mol Cancer Ther 14:111-9 (2015). PubMed: 25351918
- Jiang Q et al. MERIT40 cooperates with BRCA2 to resolve DNA interstrand cross-links. Genes Dev 29:1955-68 (2015). Human . PubMed: 26338419
- Xu W et al. TXNL1-XRCC1 pathway regulates cisplatin-induced cell death and contributes to resistance in human gastric cancer. Cell Death Dis 5:e1055 (2014). WB ; Human . PubMed: 24525731
- Luebben SW et al. A concomitant loss of dormant origins and FANCC exacerbates genome instability by impairing DNA replication fork progression. Nucleic Acids Res 42:5605-15 (2014). WB, ICC, IHC ; Mouse . PubMed: 24589582
- Schmidt L et al. ATMIN is required for the ATM-mediated signaling and recruitment of 53BP1 to DNA damage sites upon replication stress. DNA Repair (Amst) N/A:N/A (2014). PubMed: 25262557
- Lundholm L et al. Resistance to DNA-damaging treatment in non-small cell lung cancer tumor-initiating cells involves reduced DNA-PK/ATM activation and diminished cell cycle arrest. Cell Death Dis 4:e478 (2013). WB ; Human . PubMed: 23370278
- Luebben SW et al. Helq acts in parallel to Fancc to suppress replication-associated genome instability. Nucleic Acids Res N/A:N/A (2013). WB ; Mouse . PubMed: 24005041
- Adelman CA et al. HELQ promotes RAD51 paralogue-dependent repair to avert germ cell loss and tumorigenesis. Nature 502:381-4 (2013). PubMed: 24005329