Anti-MTAP 抗体 [EPR6893] (ab126770)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6893] to MTAP
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-MTAP antibody [EPR6893]
MTAP 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR6893] to MTAP -
由来種
Rabbit -
特異性
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human MTAP aa 200-300. The exact sequence is proprietary.
Database link: Q13126 -
ポジティブ・コントロール
- WB: HeLa, 293T, HT29, C6, RAW 264.7, and NIH 3T3 cell lysates. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IHC-P: Human kidney, mouse kidney, and human lung carcinoma tissue.
-
特記事項
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.
製品の特性
-
製品の状態
Liquid -
保存方法
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. -
解離定数(KD 値)
KD = 2.60 x 10 -11 M Learn more about KD -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR6893 -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab126770の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
Flow Cyt (Intra) |
1/90.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 - 1/500. |
|
WB |
1/1000 - 1/10000. Detects a band of approximately 29 kDa (predicted molecular weight: 31 kDa).
|
|
IP |
1/10 - 1/100.
|
|
IHC-P | (1) |
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat For unpurified use at 1:50 - 1:100. |
ICC/IF |
1/50 - 1/250.
|
特記事項 |
---|
Flow Cyt (Intra)
1/90. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 - 1/500. |
WB
1/1000 - 1/10000. Detects a band of approximately 29 kDa (predicted molecular weight: 31 kDa). |
IP
1/10 - 1/100. |
IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat For unpurified use at 1:50 - 1:100. |
ICC/IF
1/50 - 1/250. |
ターゲット情報
-
機能
Plays a major role in polyamine metabolism and is important for the salvage of both adenine and methionine. -
組織特異性
Ubiquitously expressed. -
配列類似性
Belongs to the PNP/MTAP phosphorylase family. -
細胞内局在
Cytoplasm. - Information by UniProt
-
参照データベース
- 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
-
別名
- 5' methylthioadenosine phosphorylase antibody
- 5''-methylthioadenosine phosphorylase antibody
- BDMF antibody
see all
画像
-
All lanes : Anti-MTAP antibody [EPR6893] (ab126770) 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?anes 1- 4: Merged signal (red and green). Green - ab126770 observed at 32 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab126770 Anti-MTAP antibody [EPR6893] 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. ab126770 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.
-
All lanes : Anti-MTAP antibody [EPR6893] (ab126770) at 1/10000 dilution (purified)
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysates
Lane 3 : Mouse kidney lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 31 kDa
Blocking and diluting buffer : 5% NFDM/TBST -
Formalin-fixed, paraffin-embedded human kidney tissue stained for MTAP with unpurified ab126770 (1/50 dilution) in immunohistochemical analysis.
Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling MTAP with Purified ab126770 at 1:1000 dilution (0.89 µg/ml). Heat mediated antigen retrieval was performed 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.
-
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling MTAP with Purified ab126770 at 1/250 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
-
ab126770 (purified) at 1:50 dilution (2µg) immunoprecipitating MTAP in HT-29 whole cell lysate.
Lane 1 (input): HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab126770 & HT-29 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126770 in HT-29 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling MTAP with purified ab126770 at 1/90 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
-
All lanes : Anti-MTAP antibody [EPR6893] (ab126770) at 1/2000 dilution (purified)
Lane 1 : HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
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: 31 kDa
Blocking and diluting buffer : 5% NFDM/TBST -
All lanes : Anti-MTAP antibody [EPR6893] (ab126770) at 1/1000 dilution (unpurified)
Lane 1 : 293T cell lysates
Lane 2 : HT29 cell lysates
Lane 3 : C6 cell lysates
Lane 4 : RAW 264.7 cell lysates
Lane 5 : NIH 3T3 cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Predicted band size: 31 kDa -
Overlay histogram showing HeLa cells stained with unpurified ab126770 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab126770, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
プロトコール
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (8)
ab126770 は 8 報の論文で使用されています。
- Aoki Y et al. Deletion of MTAP Highly Sensitizes Osteosarcoma Cells to Methionine Restriction With Recombinant Methioninase. Cancer Genomics Proteomics 19:299-304 (2022). PubMed: 35430564
- Yamamoto J et al. Linkage of methionine addiction, histone lysine hypermethylation, and malignancy. iScience 25:104162 (2022). PubMed: 35434545
- Wang T et al. Delivery of miR-654-5p via SonoVue Microbubble Ultrasound Inhibits Proliferation, Migration, and Invasion of Vascular Smooth Muscle Cells and Arterial Thrombosis and Stenosis through Targeting TCF21. Oxid Med Cell Longev 2022:4757081 (2022). PubMed: 35910838
- Abstracts from USCAP 2021: Pulmonary, Mediastinal, Pleural, and Peritoneal Pathology (910-952). Mod Pathol 34:1237-1291 (2021). PubMed: 33707674
- Abstracts from USCAP 2021: Pulmonary, Mediastinal, Pleural, and Peritoneal Pathology (910-952). Lab Invest 101:1131-1185 (2021). PubMed: 33707730
- Calvo-Vidal MN et al. Oncogenic HSP90 Facilitates Metabolic Alterations in Aggressive B-cell Lymphomas. Cancer Res 81:5202-5216 (2021). PubMed: 34479963
- Driehuis E et al. Pancreatic cancer organoids recapitulate disease and allow personalized drug screening. Proc Natl Acad Sci U S A N/A:N/A (2019). PubMed: 31818951
- Cheng XY et al. Deletion and downregulation of MTAP contribute to the motility of esophageal squamous carcinoma cells. Onco Targets Ther 10:5855-5862 (2017). PubMed: 29270023