重组Anti-MTAP抗体[EPR22570-76] (ab254265)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22570-76] to MTAP
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-MTAP抗体[EPR22570-76]
参阅全部 MTAP 一抗 -
描述
兔单克隆抗体[EPR22570-76] to MTAP -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
不适用于: IP -
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: Human liver and placenta lysates; mouse skin and liver lysates; rat skin and liver lysates; RAW 264.7, NIH/3T3, HT-29, HepG2, HeLA, Raji and Caco-2 whole cell lysates. IHC-P: Human breast and bladder cancer tissue; mouse liver tissue; rat liver tissue. ICC/IF: HT-29 cells. Flow Cyt (intra): HT-29 cells.
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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
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR22570-76 -
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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Immunohistochemistry reagents
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab254265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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Flow Cyt (Intra) |
1/400.
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WB |
1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 31 kDa).
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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说明 |
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Flow Cyt (Intra)
1/400. |
WB
1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 31 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
靶标
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功能
Plays a major role in polyamine metabolism and is important for the salvage of both adenine and methionine. -
组织特异性
Ubiquitously expressed. -
序列相似性
Belongs to the PNP/MTAP phosphorylase family. -
细胞定位
Cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 4507 Human
- Entrez Gene: 66902 Mouse
- Entrez Gene: 298227 Rat
- Omim: 156540 Human
- SwissProt: Q13126 Human
- SwissProt: Q9CQ65 Mouse
- Unigene: 193268 Human
- Unigene: 28500 Mouse
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别名
- 5' methylthioadenosine phosphorylase antibody
- 5''-methylthioadenosine phosphorylase antibody
- BDMF antibody
see all
图片
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All lanes : Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MTAP knockout HeLa cell lysate
Lane 3 : HT-29 cell lysate
Lane 4 : A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab254265 observed at 32 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab254265 Anti-MTAP antibody [EPR22570-76] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265272 (knockout cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab254265 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution
Lane 1 : Mouse skin lysate
Lane 2 : Mouse liver lysate
Lane 3 : Rat skin lysate
Lane 4 : Rat liver lysate
Lane 5 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 6 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 31 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times:
Lanes 1-4: 15 seconds; Lane 5: 10 seconds; Lane 6: 3.25 seconds.The expression molecular weight observed is consistent with what has been described in the literature (PMID:15492751).
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All lanes : Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution
Lane 1 : Human liver lysate
Lane 2 : Human placenta lysate
Lane 3 : HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate
Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 5 : HeLa (human epithelial cell line from cervix adenocarcinoma0 whole cell lysate
Lane 6 : Raji 9human Burkitt's lymphoma cell line) whole cell lysate
Lane 7 : Caco-2 (human colorectal adenocarcinoma cell line) whole cell lysate
Lane 8 : A549 9human colorectal adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 3-8 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 31 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Negative control: A549 (PMID: 17548352).
The expression molecular weight observed is consistent with what has been described in the literature (PMID:15492751).
Exposure times.
Lanes 1-2: 10 seconds; Lane3: 5.5 seconds; Lanes 4-8: 3.25 seconds.
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Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) (Left) anf H-29 (human colorectal adenocarcinoma cell line) (Right) cells labeling MTAP with ab254265 at 1/400 dilution (red) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.
Negative control: A549 (PMID: 8971171.
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Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in rat liver (PMID:15492751) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in mouse liver (PMID:15492751) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in stroma cells and no staining in tumor cells of human bladder cancer (PMID:27270441) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human breast tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in human breast (PMID:26751376, 18712977) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) and HT-29 (human colorectal adenocarcinoma cell line) cells labeling MTAP with ab254265 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in HT-29 cell line. The nuclear counter stain is DAPI (blue) Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
SEcondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
Negative control: A549(PMID: 8971171.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (0)
ab254265 尚未被引用在任何文献中。