重组Anti-Paxillin抗体[Y113] (ab32084)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y113] to Paxillin
- Suitable for: WB, ICC/IF, Flow Cyt (Intra), IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-Paxillin抗体[Y113]
参阅全部 Paxillin 一抗 -
描述
兔单克隆抗体[Y113] to Paxillin -
宿主
Rabbit -
特异性
This antibody recognises Paxillin alpha, beta and gamma isoforms.
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经测试应用
适用于: WB, ICC/IF, Flow Cyt (Intra), IP, IHC-Pmore details -
种属反应性
与反应: Mouse, Human
预测可用于: Dog -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- Flow Cyt (intra): HeLa cells. ICC/IF: HeLa cells IP: HeLa whole cell lysate WB: HeLa, RAW 264.7 and mouse heart lysate. IHC-P: Human cerebrum tissue and human breast carcinoma tissue.
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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. -
解离常数(KD)
KD = 4.17 x 10 -10 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y113 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- HRP Anti-Paxillin antibody [Y113] (ab197612)
- Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
- Alexa Fluor® 488 Anti-Paxillin antibody [Y113] (ab246718)
- Alexa Fluor® 647 Anti-Paxillin antibody [Y113] (ab246719)
- APC Anti-Paxillin antibody [Y113] (ab319338)
- PE Anti-Paxillin antibody [Y113] (ab319473)
- Alexa Fluor® 594 Anti-Paxillin antibody [Y113] (ab319831)
- Alexa Fluor® 555 Anti-Paxillin antibody [Y113] (ab319972)
- Alexa Fluor® 750 Anti-Paxillin antibody [Y113] (ab321569)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Related Products
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Substrate reagent
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32084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 | (10) |
1/1000. Predicted molecular weight: 68 kDa.
For unpurified use at 1/5000 - 1/10000. |
ICC/IF | (7) |
1/50.
For unpurified use at 1/250. |
Flow Cyt (Intra) |
1/100.
|
|
IP | (1) |
1/20.
|
IHC-P | (1) |
1/1200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
说明 |
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WB
1/1000. Predicted molecular weight: 68 kDa. For unpurified use at 1/5000 - 1/10000. |
ICC/IF
1/50. For unpurified use at 1/250. |
Flow Cyt (Intra)
1/100. |
IP
1/20. |
IHC-P
1/1200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
靶标
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功能
Cytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion). -
序列相似性
Belongs to the paxillin family.
Contains 4 LIM zinc-binding domains. -
翻译后修饰
Phosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens. -
细胞定位
Cytoplasm > cytoskeleton. Cell junction > focal adhesion. - Information by UniProt
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数据库链接
- Entrez Gene: 5829 Human
- Entrez Gene: 19303 Mouse
- Omim: 602505 Human
- SwissProt: P49023 Human
- SwissProt: Q8VI36 Mouse
- Unigene: 446336 Human
- Unigene: 18714 Mouse
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别名
- FLJ16691 antibody
- FLJ23042 antibody
- Paired box protein Pax 1 antibody
see all
图片
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All lanes : Anti-Paxillin antibody [Y113] (ab32084) at 1/1000 dilution
Lane 1 : Wild-type A431 cell lysate
Lane 2 : PXN knockout A431 cell lysate
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Anti-PXN antibody [Y113] (ab32084) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32084 was shown to bind specifically to PXN. A band was observed at 70 kDa in wild-type A431 cell lysates with no signal observed at this size in PXN knockout cell line ab261892 (knockout cell lysate ab261701). To generate this image, wild-type and PXN knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween$®$ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
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All lanes : Anti-Paxillin antibody [Y113] (ab32084) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : Mouse heart lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 68 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] (ab32084)
Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue sections labelling Paxillin with ab32084 at 1/1200 dilution. The section was incubated with ab32084 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Positive staining on human breast carcinoma tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Paxillin with purified ab32084 at 1/50 dilution (2.88 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.
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Intracellular Flow Cytometry analysis ofHeLa (human cervix adenocarcinoma) cells labeling with purified ab32084 at 1/100 dilution (10µg/ml) (Red). Cells were fixed with4% paraformaldehydeand permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody.Rabbit monoclonal IgG (Black) (ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] (ab32084)
Immunohistochemistry analysis of paraffin-embedded human cerebrum tissue sections labelling Paxillin with ab32084 at 1/1200 dilution. The section was incubated with ab32084 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Positive staining on human cerebrum tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Purified ab32084 at 1/20 dilution (0.7µg) immunoprecipitating Paxillin in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg.
Lane 2 (+): ab32084 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32084 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 48 kDa
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
文献 (214)
ab32084 被引用在 214 文献中.
- Sonam S et al. Mechanical stress driven by rigidity sensing governs epithelial stability. Nat Phys 19:132-141 (2023). PubMed: 36686215
- Tiskratok W et al. Substrate stiffness controls proinflammatory responses in human gingival fibroblasts. Sci Rep 13:1358 (2023). PubMed: 36693942
- Miyauchi M et al. BAG6 supports stress fiber formation by preventing the ubiquitin-mediated degradation of RhoA. Mol Biol Cell 34:ar34 (2023). PubMed: 36884293
- Lu S et al. CD248 promotes migration and metastasis of osteosarcoma through ITGB1-mediated FAK-paxillin pathway activation. BMC Cancer 23:290 (2023). PubMed: 36997926
- Melamed S et al. Initiation of fibronectin fibrillogenesis is an enzyme-dependent process. Cell Rep 42:112473 (2023). PubMed: 37148241