重组Anti-53BP1抗体[EPR2172(2)] (ab175933)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2172(2)] to 53BP1
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
-
产品名称
Anti-53BP1抗体[EPR2172(2)]
参阅全部 53BP1 一抗 -
描述
兔单克隆抗体[EPR2172(2)] to 53BP1 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, ICC/IF, 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.
-
阳性对照
- WB: HepG2 and HeLa cell lysate and human fetal heart and fetal brain tissue lysates, mouse heart and rat heart tissue lysates. IHC-P: human colon, liver carcinoma and tonsil, mouse and rat liver tissues. ICC/IF: HepG2 cells.
-
常规说明
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
-
形式
Liquid -
存放说明
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. -
存储溶液
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR2172(2) -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab175933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 |
---|---|---|
WB |
1/1000 - 1/5000. Detects a band of approximately 450 kDa (predicted molecular weight: 214 kDa).
|
|
IHC-P |
1/60 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
|
ICC/IF | (1) |
1/100 - 1/250.
|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
说明 |
---|
WB
1/1000 - 1/5000. Detects a band of approximately 450 kDa (predicted molecular weight: 214 kDa). |
IHC-P
1/60 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
靶标
-
功能
May have a role in checkpoint signaling during mitosis. Enhances TP53-mediated transcriptional activation. Plays a role in the response to DNA damage. -
疾病相关
Note=A chromosomal aberration involving TP53BP1 is found in a form of myeloproliferative disorder chronic with eosinophilia. Translocation t(5;15)(q33;q22) with PDGFRB creating a TP53BP1-PDGFRB fusion protein. -
序列相似性
Contains 2 BRCT domains. -
翻译后修饰
Asymmetrically dimethylated on Arg residues by PRMT1. Methylation is required for DNA binding.
Phosphorylated at basal level in the absence of DNA damage. Hyper-phosphorylated in an ATM-dependent manner in response to DNA damage induced by ionizing radiation. Hyper-phosphorylated in an ATR-dependent manner in response to DNA damage induced by UV irradiation. -
细胞定位
Nucleus. Chromosome > centromere > kinetochore. Associated with kinetochores. Both nuclear and cytoplasmic in some cells. Recruited to sites of DNA damage, such as double stand breaks. Methylation of histone H4 at 'Lys-20' is required for efficient localization to double strand breaks. - Information by UniProt
-
数据库链接
- Entrez Gene: 7158 Human
- Entrez Gene: 27223 Mouse
- Entrez Gene: 296099 Rat
- Omim: 605230 Human
- SwissProt: Q12888 Human
- SwissProt: P70399 Mouse
- Unigene: 440968 Human
- Unigene: 383499 Mouse
see all -
别名
- 53 BP1 antibody
- 53BP1 antibody
- FLJ41424 antibody
see all
图片
-
Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling 53BP1 with purified ab175933 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/200) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
-
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: 53BP1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (40 µg)
Lane 4: HepG2 cell lysate (40 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab175933 observed at 350 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab175933 was shown to specifically react with 53BP1 when 53BP1 knockout samples were used. Wild-type and 53BP1 knockout samples were subjected to SDS-PAGE. ab175933 and ab18058 (loading control to Vinculin) were diluted 1/1000 and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue labelling 53BP1 with purified ab175933 at a dilution of 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
-
ab175933 staining 53BP1in the human cell lineHepG2 (human hepatocellular carcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/30. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
-
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: 53BP1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (40 µg)
Lane 4: HepG2 cell lysate (40 µg)Lanes 1 - 4: Merged signal (red and green).
Green - Target observed at 350 kDa. Red - loading control, ab18058, observed at 124 kDa.
This western blot image is a comparison between ab175933 and a competitor's top cited rabbit polyclonal antibody.
-
All lanes : Anti-53BP1 antibody [EPR2172(2)] (ab175933) at 1/1000 dilution (purified)
Lane 1 : Mouse heart tissue lysate
Lane 2 : Mouse brain tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 214 kDa
Observed band size: 450 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM /TBST.
-
Anti-53BP1 antibody [EPR2172(2)] (ab175933) at 20 µg (purified) + Rat heart tissue lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 214 kDa
Observed band size: 450 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM /TBST.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling 53BP1 with purified ab175933 at a dilution of 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
-
All lanes : Anti-53BP1 antibody [EPR2172(2)] (ab175933) at 1/5000 dilution (purified)
Lane 1 : Human fetal heart tissue lysate
Lane 2 : HeLa whole cell lysate
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: 214 kDa
Observed band size: 450 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM /TBST.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver carcinoma tissue labelling 53BP1 with purified ab175933 at a dilution of 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
-
All lanes : Anti-53BP1 antibody [EPR2172(2)] (ab175933) at 1/1000 dilution (unpurified)
Lane 1 : HepG2 cell lysate
Lane 2 : Human fetal brain lysate
Lane 3 : Human fetal heart lysate
Lysates/proteins at 1/10 dilution per lane.
Predicted band size: 214 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling 53BP1 with unpurified ab175933 at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling 53BP1 with unpurified ab175933 at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling 53BP1 with unpurified ab175933 at a dilution of 1/100.
实验方案
数据表及文件
-
SDS download
-
Datasheet download
文献 (39)
ab175933 被引用在 39 文献中.
- Coleman KE et al. USP1-trapping lesions as a source of DNA replication stress and genomic instability. Nat Commun 13:1740 (2022). PubMed: 35365626
- Zhang J et al. Flap endonuclease 1 and DNA-PKcs synergistically participate in stabilizing replication fork to encounter replication stress in glioma cells. J Exp Clin Cancer Res 41:140 (2022). PubMed: 35414100
- Lu H et al. DNA-PKcs-dependent phosphorylation of RECQL4 promotes NHEJ by stabilizing the NHEJ machinery at DNA double-strand breaks. Nucleic Acids Res 50:5635-5651 (2022). PubMed: 35580045
- Tang Y et al. Pressure Loading Induces DNA Damage in Human Hepatocyte Line L02 Cells via the ERK1/2-Dicer Signaling Pathway. Int J Mol Sci 23:N/A (2022). PubMed: 35628153
- Li M et al. METTL3 antagonizes 5‑FU chemotherapy and confers drug resistance in colorectal carcinoma. Int J Oncol 61:N/A (2022). PubMed: 35856434