重组Anti-PARP1抗体[E102] - BSA and Azide free (ab221923)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E102] to PARP1 - BSA and Azide free
- Suitable for: IHC-P, WB, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-PARP1抗体[E102] - BSA and Azide free
参阅全部 PARP1 一抗 -
描述
兔单克隆抗体[E102] to PARP1 - BSA and Azide free -
宿主
Rabbit -
特异性
This antibody recognises both pro-form and p25 cleaved form of PARP1.
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经测试应用
适用于: IHC-P, WB, ICC/IF, Flow Cyt (Intra)more details
不适用于: IP -
种属反应性
与反应: 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: Wild type HAP1 whole cell lysate; HeLa whole cell lysate (ab150035); HEK-293T cell lysate; ICC/IF: HeLa cells; IHC-P: Human breast carcinoma tissue; Flow Cyt (intra): HeLa cells, HAP1 cells. WB: Jurkat whole cell lysate.
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常规说明
Ab221923 is the carrier-free version of ab32138..
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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
E102 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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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ab221923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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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 113 kDa.
Existing as a 113 kDa nuclear protein, PARP1 is cleaved between amino acids Asp214 and Gly215 to yield two fragments of 29 kDa (C-terminal catalytic domain) and 85 kDa (N-terminal DNA-binding domain) |
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376- Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 113 kDa. Existing as a 113 kDa nuclear protein, PARP1 is cleaved between amino acids Asp214 and Gly215 to yield two fragments of 29 kDa (C-terminal catalytic domain) and 85 kDa (N-terminal DNA-binding domain) |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376- Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
Involved in the base excision repair (BER) pathway, by catalyzing the poly(ADP-ribosyl)ation of a limited number of acceptor proteins involved in chromatin architecture and in DNA metabolism. This modification follows DNA damages and appears as an obligatory step in a detection/signaling pathway leading to the reparation of DNA strand breaks. Mediates the poly(ADP-ribosyl)ation of APLF and CHFR. Positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150. -
序列相似性
Contains 1 BRCT domain.
Contains 1 PARP alpha-helical domain.
Contains 1 PARP catalytic domain.
Contains 2 PARP-type zinc fingers. -
翻译后修饰
Phosphorylated by PRKDC. Phosphorylated upon DNA damage, probably by ATM or ATR.
Poly-ADP-ribosylated by PARP2. Poly-ADP-ribosylation mediates the recruitment of CHD1L to DNA damage sites.
S-nitrosylated, leading to inhibit transcription regulation activity. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 142 Human
- Omim: 173870 Human
- SwissProt: P09874 Human
- Unigene: 177766 Human
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别名
- ADP ribosyltransferase (NAD+; poly (ADP ribose) polymerase) antibody
- ADP ribosyltransferase antibody
- ADP ribosyltransferase diphtheria toxin like 1 antibody
see all
图片
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32138).
Flow cytometry overlay histogram showing wild-type Hap1 (green line) and PARP1 knockout Hap1 stained with ab32138 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab32138) (1x 106 in 100μl at 0.04 μg/ml (1/55750)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type Hap1 - black line, PARP1 knockout Hap1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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All lanes : Anti-PARP1 antibody [E102] (ab32138) at 1/1000 dilution (Purified)
Lane 1 : Untreated Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) treated with 1µM staurosporine for 4 hours whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 113 kDapro-form: 116kDa; p25 caspases cleaved form: 25kDa; proteolysis cleaved fragments: 58kDa and 42kDa
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All lanes : Anti-PARP1 antibody [E102] (ab32138) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : PARP1 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 113 kDa
Observed band size: 113 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32138).
Lanes 1- 2: Merged signal (red and green). Green - ab32138 observed at 113 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32138 was shown to react with PARP1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266598 (knockout cell lysate ab257017). Wild-type HEK-293T and PARP1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab32138 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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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This data was developed using ab32138, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling PARP1 with purified ab32138 at 1/200 dilution (0.51 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. -
This data was developed using ab32138, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PARP1 with purified ab32138 at 1/100 dilution (1.0 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) 1/200 (2.5 µg/mL). 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 ab32138, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling PARP1 with purified ab32138 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
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