Anti-LAG-3 抗体 [CAL26] (ab237719)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [CAL26] to LAG-3
- Suitable for: WB, IP, Flow Cyt (Intra), IHC-P
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-LAG-3 antibody [CAL26]
LAG-3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [CAL26] to LAG-3 -
由来種
Rabbit -
アプリケーション
適用あり: WB, IP, Flow Cyt (Intra), IHC-Pmore details
適用なし: ICC/IF -
種交差性
交差種: 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: LAG-3-GFP transfected HEK-293 whole cell lysate; human PBMC whole cell lysate. IHC-P: Human tonsil tissue. Flow cyt: Human PBMCs treated with 10µg/ml PHA for 48 hours. IP: LAG-3-GFP transfected HEK-203& whole cell lysate.
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.50
Preservative: 0.05% Sodium azide
Constituent: PBS -
Concentration information loading...
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精製度
Protein A purified -
特記事項(精製)
Purity >99%. -
ポリ/モノ
モノクローナル -
クローン名
CAL26 -
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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Immunohistochemistry reagents
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Isotype control
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ab237719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 |
1/1000. Detects a band of approximately 16, 70 kDa (predicted molecular weight: 57 kDa).
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IP |
1/20.
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Flow Cyt (Intra) |
1/250.
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IHC-P |
Use a concentration of 0.3 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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特記事項 |
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WB
1/1000. Detects a band of approximately 16, 70 kDa (predicted molecular weight: 57 kDa). |
IP
1/20. |
Flow Cyt (Intra)
1/250. |
IHC-P
Use a concentration of 0.3 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ターゲット情報
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機能
Involved in lymphocyte activation. Binds to HLA class-II antigens. -
組織特異性
On cell surface of activated NK and T-lymphocytes. -
配列類似性
Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
細胞内局在
Membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 3902 Human
- Omim: 153337 Human
- SwissProt: P18627 Human
- Unigene: 409523 Human
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別名
- CD223 antibody
- CD223 antigen antibody
- FDC protein antibody
see all
画像
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All lanes : Anti-LAG-3 antibody [CAL26] (ab237719) at 1/1000 dilution
Lane 1 : HDLM-2 cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 57,70 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-LAG-3 antibody [CAL26] 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, ab237719 was shown to bind specifically to LAG-3. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling LAG-3 with ab237719 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the human tonsil 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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LAG-3 was immunoprecipitated from 0.35 mg of LAG-3-GFP transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate with ab237719 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab237719 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.
Lane 1: LAG-3-GFP transfected HEK-293T whole cell lysate 10 μg (Input).
Lane 2: ab237719 IP in LAG-3-GFP transfected HEK-293T whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237719 in LAG-3-GFP transfected HEK-293T whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds. -
Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized human peripheral blood mononuclear cells ((PBMC) (treated with 10μg/ml PHA for 48 hours) cell line labeling LAG-3 with ab237719 at 1/250 dilution (Right) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left).
Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed (ab150097), at 1/5000 dilution was used as the secondary antibody.
Cells were surface stained with anti-CD3 conjugated to Alexa Fluor® 647. Then fixed with 2% PFA for 10min followed by intracellular staining rabbit IgG (Left) or ab237719 (Right).
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All lanes : Anti-LAG-3 antibody [CAL26] (ab237719) at 1/1000 dilution
Lane 1 : Untransfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : LAG-3-GFP overexpression HEK-293T 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: 57 kDa
Observed band size: 16,70 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and dilution buffer: 5% NFDM/TBST.
This antibody also detects a 16kDa cleaved fragment. (PMID: 15557174).
This blot was developed using a higher sensitivity ECL substrate.
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All lanes : Anti-LAG-3 antibody [CAL26] (ab237719) at 1/1000 dilution
Lane 1 : Human peripheral blood mononuclear cell (PBMC), whole cell lysate
Lane 2 : Human peripheral blood mononuclear cell (PBMC) treated with 10µg/ml PHA for 48 hours, whole cell lysate
Lane 3 : Human lymphoma 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: 57 kDa
Observed band size: 16,70 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and dilution buffer: 5% NFDM/TBST.
This antibody also detects a 16kDa cleaved fragment. (PMID: 15557174).
This blot was developed using a higher sensitivity ECL substrate.
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Formalin-fixed, paraffin-embedded human tonsil tissue stained for LAG-3 using ab237719 at 0.3 μg/ml in immunohistochemical analysis.
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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SDS download
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Datasheet download
参考文献 (0)
ab237719 は論文での使用が確認できていません。