Anti-PD-L1 抗体 [CAL10] - Mouse IgG1 (Chimeric) (ab279292)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [CAL10] to PD-L1 - Mouse IgG1
- Suitable for: WB, Flow Cyt (Intra), IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-PD-L1 antibody [CAL10] - Mouse IgG1 (Chimeric)
PD-L1 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [CAL10] to PD-L1 - Mouse IgG1 -
由来種
Mouse -
アプリケーション
適用あり: WB, Flow Cyt (Intra), IHC-P, ICC/IFmore details -
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: PD-L1 stably expressed CHO whole cell lysate. Human placenta tissue lysate. NCI-H1299 whole cell lysate. ICC: PD-L1 stably expressed CHO cells. Flow Cyt (intra): PD-L1 stably expressed CHO cells. IHC-P: Human tonsil tissue.
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特記事項
This mouse monoclonal chimeric antibody has been engineered from a RabMAb parent antibody (ab237726). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.
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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
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
CAL10 -
アイソタイプ
IgG1 -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Related Products
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ab279292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) |
1/1000.
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Flow Cyt (Intra) |
1/50.
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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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ICC/IF |
1/100.
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特記事項 |
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WB
1/1000. |
Flow Cyt (Intra)
1/50. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
ターゲット情報
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機能
Involved in the costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNG, in an IL2-dependent and a PDCD1-independent manner. Interaction with PDCD1 inhibits T-cell proliferation and cytokine production. -
組織特異性
Highly expressed in the heart, skeletal muscle, placenta and lung. Weakly expressed in the thymus, spleen, kidney and liver. Expressed on activated T- and B-cells, dendritic cells, keratinocytes and monocytes. -
配列類似性
Belongs to the immunoglobulin superfamily. BTN/MOG family.
Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
細胞内局在
Cell membrane and Endomembrane system. - Information by UniProt
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参照データベース
- Entrez Gene: 29126 Human
- Omim: 605402 Human
- SwissProt: Q9NZQ7 Human
- Unigene: 521989 Human
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別名
- B7 H antibody
- B7 H1 antibody
- B7 homolog 1 antibody
see all
画像
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All lanes : Anti-PD-L1 antibody [CAL10] - Mouse IgG1 (Chimeric) (ab279292) at 1/1000 dilution
Lane 1 : Wild-type A549 Control IFN-gamma (0 ng/mL, 48 h), ab255450
Lane 2 : Wild-type A549 Treated IFN-gamma (100 ng/mL, 48 h), ab255450
Lane 3 : CD274 knockout A549 Control IFN-gamma (0 ng/mL, 48 h), ab267055
Lane 4 : CD274 knockout A549 Treated IFN-gamma (100 ng/mL, 48 h), ab267055
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 45 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-PD-L1 antibody [CAL10] – Mouse IgG1 (Chimeric) staining at 1/1000 dilution, shown in green; Rabbit anti-Vinculin antibody (ab219649) loading control staining at 1/1000 dilution, shown in red. In Western blot, ab279292 was shown to bind specifically to PD-L1. A band was observed at 45 kDa in wild-type A549 cell lysates with no signal observed at this size in Cd274 knockout cell line. To generate this image, wild-type and Cd274 knockout A549 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-PD-L1 antibody [CAL10] - Mouse IgG1 (Chimeric) (ab279292) at 1/1000 dilution
Lane 1 : Wild-type A549 Treated IFN-gamma (100 ng/ml) for 48 hours cell lysate
Lane 2 : CD274 knockout A549 Treated IFN-gamma (100 ng/ml) for 48 hours cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 48 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-PD-L1 antibody [CAL10] – Mouse IgG1 (Chimeric) staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab279292 was shown to bind specifically to PD-L1. A band was observed at 48 kDa in treated wild-type A549 cell lysates with no signal observed at this size in Cd274 knockout cell line ab267054 (knockout cell lysate ab256831). To generate this image, wild-type and Cd274 knockout A549 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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All lanes : Anti-PD-L1 antibody [CAL10] - Mouse IgG1 (Chimeric) (ab279292) at 1/1000 dilution
Lane 1 : CHO-S (Chinese hamster ovary epithelial cell) whole cell lysate
Lane 2 : CHO-PD-L1 (PD-L1 stably expressed Chinese hamster ovary epithelial cell) whole cell lysate
Lane 3 : Human placenta tissue lysate
Lane 4 : NCI-H1299 (human lung carcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution
Observed band size: 40-60 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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IHC image of PD-L1 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab279292, 1ug/ml, for 15 mins at room temperature. A rabbit anti-mouse IgG1, ab125913, was added for 8 mins at room temperature and detected using an HRP conjugated goat anti-rabbit compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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Immunocytochemical analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100-fixed permeabilized CHO-PD-L1 cells labeling PD-L1 with ab279292 at 1/100 dilution, followed by ab150113 Goat Anti-MouseIgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/500 dilution (Red). The nuclear counterstain was DAPI (Blue). Confocal image showing membranous and cytoplasmic staining in CHO-PD-L1 cells.
Negative control 1: ab279292 at a 1/100 dilution followed by ab150080 at a 1/200 dilution.
Negative control 2: ab179513 at a 1/200 dilution followed by ab150157 at a 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized CHO-s (Chinese hamster ovary epithelial cell, Blue) / CHO-PDL1 (PD-L1 stably expessed Chinese hamster ovary epithelial cell, Red) labelling PD-L1 with ab279292 at 1/50 dilution (0.1µg).
Goat Anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
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)
ab279292 は論文での使用が確認できていません。