Anti-CrkL 抗体 [Y244] - BSA and Azide free (ab247210)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y244] to CrkL - BSA and Azide free
- Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-CrkL antibody [Y244] - BSA and Azide free
CrkL 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [Y244] to CrkL - BSA and Azide free -
由来種
Rabbit -
特異性
This antibody recognises CrkL (Crk-like)protein.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat
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アプリケーション
適用あり: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
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
- WB: Wild-type HeLa, CRKL knockout HeLa and K-562 cells. K-562 whole cell lysate. Mouse and rat thymus lysate. ICC/IF: K-562 cells; IHC-P: Human colon tissue; Flow Cyt: K-562 cells.
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特記事項
ab247210 is the carrier-free version of ab32018.
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. -
バッファー
pH: 7.2
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
Y244 -
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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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
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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ab247210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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WB |
Use at an assay dependent concentration. Detects a band of approximately 36 kDa (predicted molecular weight: 33 kDa).
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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.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat |
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Flow Cyt |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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特記事項 |
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WB
Use at an assay dependent concentration. Detects a band of approximately 36 kDa (predicted molecular weight: 33 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 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 |
Flow Cyt
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
ターゲット情報
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機能
May mediate the transduction of intracellular signals. -
配列類似性
Belongs to the CRK family.
Contains 1 SH2 domain.
Contains 2 SH3 domains. - Information by UniProt
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参照データベース
- Entrez Gene: 1399 Human
- Entrez Gene: 12929 Mouse
- Entrez Gene: 287942 Rat
- Omim: 602007 Human
- SwissProt: P46109 Human
- SwissProt: P47941 Mouse
- SwissProt: Q5U2U2 Rat
- Unigene: 5613 Human
see all -
別名
- Crk L antibody
- Crk like protein antibody
- Crk-like protein antibody
see all
画像
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All lanes : Anti-CrkL antibody [Y244] (ab32018) at 1/1000 dilution (Purified)
Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 2 : Mouse thymus lysate
Lane 3 : Rat thymus lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 33 kDa -
All lanes : Anti-CrkL antibody [Y244] (ab32018) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CRKL knockout HeLa cell lysate
Lane 3 : K-562 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 33 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab32018).
Lanes 1-3: Merged signal (red and green). Green - ab32018 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
ab32018 Anti-CrkL antibody [Y244] was shown to specifically react with CrkL in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265993 (knockout cell lysate ab257397) was used. Wild-type and CrkL knockout samples were subjected to SDS-PAGE. ab32018 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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 20000 dilution for 1 hour at room temperature before imaging.
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This data was developed using ab32018, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling CrkL with purified ab32018 at 1/100 dilution (2.24 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
This data was developed using ab32018, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling CrkL with purified ab32018 at 1/50 dilution (4.5 µg/mL). Cells were fixed in 100% Methanol 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 ab32018, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labelling CrkL with purified ab32018 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). -
This data was developed using ab32018, the same antibody clone in a different buffer formulation.
Purified ab32018 at 1/50 dilution (2µg) immunoprecipitating CrkL in K-562 whole cell lysate.
Lane 1 (input): K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg
Lane 2 (+): ab32018 + K-562 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32018 in K-562 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: 37 kDa
プロトコール
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.
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (0)
ab247210 は論文での使用が確認できていません。