Anti-Paxillin 抗体 [Y113] - BSA and Azide free (ab216652)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Paxillin antibody [Y113] - BSA and Azide free
Paxillin 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [Y113] to Paxillin - BSA and Azide free -
由来種
Rabbit -
特異性
This antibody recognises Paxillin alpha, beta and gamma isoforms.
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アプリケーション
適用あり: Flow Cyt (Intra), ICC/IF, IHC-P, WB, IPmore details -
種交差性
交差種: Mouse, Human
交差が予測される動物種: Dog -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- 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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特記事項
ab216652 is the carrier-free version of ab32084.
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
解離定数(KD 値)
KD = 4.17 x 10 -10 M Learn more about KD -
バッファー
pH: 7.20
Constituent: PBS -
キャリア・フリー
はい -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
Y113 -
アイソタイプ
IgG -
研究分野
関連製品
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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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Positive Controls
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Related Products
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Substrate reagent
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ab216652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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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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: 68 kDa.
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IP |
Use at an assay dependent concentration.
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特記事項 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
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: 68 kDa. |
IP
Use at an assay dependent concentration. |
ターゲット情報
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機能
Cytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion). -
配列類似性
Belongs to the paxillin family.
Contains 4 LIM zinc-binding domains. -
翻訳後修飾
Phosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens. -
細胞内局在
Cytoplasm > cytoskeleton. Cell junction > focal adhesion. - Information by UniProt
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参照データベース
- 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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別名
- FLJ16691 antibody
- FLJ23042 antibody
- Paired box protein Pax 1 antibody
see all
画像
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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.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?This data was developed using ab32084, the same antibody clone in a different buffer formulation.
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 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.
This data was developed using ab32084, the same antibody clone in a different buffer formulation. -
This data was developed using ab32084, the same antibody clone in a different buffer formulation.
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. -
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling with purified ab32084 at 1/100 dilution (10ug/ml) (Red). Cells were fixed with 4% paraformaldehyde and 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).
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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.
This data was developed using ab32084, the same antibody clone in a different buffer formulation. -
This data was developed using ab32084, the same antibody clone in a different buffer formulation.
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. -
ab32084 (Unpurified format) showing positive staining in Normal ovary tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab32084 (Unpurified format) showing positive staining in Ovarian carcinoma tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab32084 (Unpurified format) showing positive staining in Transitional cell carcinoma of kidney tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).
プロトコール
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (14)
ab216652 は 14 報の論文で使用されています。
- Lin LL et al. PAI-1-Dependent Inactivation of SMAD4-Modulated Junction and Adhesion Complex in Obese Endometrial Cancer. Cell Rep 33:108253 (2020). PubMed: 33053339
- Yariswamy M et al. Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction. J Biol Chem 291:19425-36 (2016). PubMed: 27466370
- Izumi D et al. CXCL12/CXCR4 activation by cancer-associated fibroblasts promotes integrin ß1 clustering and invasiveness in gastric cancer. Int J Cancer 138:1207-19 (2016). ICC/IF ; Human . PubMed: 26414794
- Fuhrmann A et al. Cation type specific cell remodeling regulates attachment strength. PLoS One 9:e102424 (2014). ICC/IF ; Mouse . PubMed: 25014042
- Chen DL et al. Abnormal expression of paxillin correlates with tumor progression and poor survival in patients with gastric cancer. J Transl Med 11:277 (2013). IHC-P ; Human . PubMed: 24180516