Anti-CD47 抗体 [EPR24922-5] (BSA and Azide free) (ab300436)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24922-5] to CD47 - BSA and Azide free
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-CD47 antibody [EPR24922-5] (BSA and Azide free)
CD47 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR24922-5] to CD47 - BSA and Azide free -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-Pmore details
適用なし: Flow Cyt,ICC/IF or IP -
種交差性
交差種: Human
非交差種: Mouse, Rat -
免疫原
Recombinant fragment. 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: Jurkat, U937, NIH:OVCAR-3, Wild-type HEK-293T, CD47 CRISPR-Cas9 edited HEK-293T, whole cell lysates; IHC-P: Human tonsil, human ovarian cancer, human gastric cancer FFPE tissue sections; human wild-type HEK-293T cell pellet, human CD47 CRISPR-Cas9 edited HEK-293T cell pellet.
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特記事項
ab300436 is the carrier-free version of ab300435.
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. -
バッファー
pH: 7.20
Constituent: 100% 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
EPR24922-5 -
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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KO cell lines
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KO cell lysates
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ab300436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.
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IHC-P |
Use at an assay dependent concentration. 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
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. 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
Has a role in both cell adhesion by acting as an adhesion receptor for THBS1 on platelets, and in the modulation of integrins. Plays an important role in memory formation and synaptic plasticity in the hippocampus (By similarity). Receptor for SIRPA, binding to which prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells. Interaction with SIRPG mediates cell-cell adhesion, enhances superantigen-dependent T-cell-mediated proliferation and costimulates T-cell activation. May play a role in membrane transport and/or integrin dependent signal transduction. May prevent premature elimination of red blood cells. May be involved in membrane permeability changes induced following virus infection. -
組織特異性
Very broadly distributed on normal adult tissues, as well as ovarian tumors, being especially abundant in some epithelia and the brain. -
配列類似性
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
細胞内局在
Cell membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 961 Human
- Omim: 601028 Human
- SwissProt: Q08722 Human
- Unigene: 446414 Human
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別名
- Antigen identified by monoclonal antibody 1D8 antibody
- Antigenic surface determinant protein OA3 antibody
- CD 47 antibody
see all
画像
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All lanes : Anti-CD47 antibody [EPR24922-5] (ab300435) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate
Lane 3 : U937 (human histiocytic lymphoma monocyte) whole cell lysate
Lane 4 : NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell) 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
Observed band size: 47,52 kDa why is the actual band size different from the predicted?This data was developed using ab300435, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: HepG2 (PMID: 28378740)
Samples are non-boiled as boiling may cause protein aggregates.
Exposure time: Lane 1-3: 48 seconds Lane 4: 15 seconds
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All lanes : Anti-CD47 antibody [EPR24922-5] (ab300435) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : CD47 CRISPR-Cas9 edited HEK-293T whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Lane 4 : HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Observed band size: 47,52 kDa why is the actual band size different from the predicted?This data was developed using ab300435, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBSTIntercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Samples are non-boiled as boiling may cause protein aggregates.
False colour image of Western blot: Anti-CD47 antibody [EPR24922-5] (ab300435) 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, ab300435 was shown to bind specifically to CD47. A diffuse band was observed at 47-52 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in CD47 CRISPR-Cas9 edited cell line ab266324 (CRISPR-Cas9 edited cell lysate ab257220). The band observed in the CRISPR-Cas9 edited lysate lane above 90kDa is likely to represent CD47 with an insertion. The band below 47 kDa in the CRISPR-Cas9 edited lysate lane is likely to represent a truncated form of CD47. This has not been investigated further and the functional properties of the gene products have not been determined. To generate this image, wild-type and CD47 CRISPR-Cas9 edited HEK-293T cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS 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.
Negative control: HepG2 (PMID: 28378740)
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This data was developed using ab300435, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human wild-type and CD47 CRISPR-Cas9 edited cells labeling CD47 with ab300435 at 1/500 dilution (0.098 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining on (A) Human wild-type HEK-293T cells and almost no staining on (B) Human CD47 CRISPR-Cas9 edited HEK-293T cells (ab266324). The section was incubated with ab300435 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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This data was developed using ab300435, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling CD47 with ab300435 at 1/2000 dilution (0.244 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining on human gastric cancer (PMID: 28693236). The section was incubated with ab300435 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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This data was developed using ab300435, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling CD47 with ab300435 at 1/2000 dilution (0.244 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining on human ovarian cancer (PMID: 28380460). The section was incubated with ab300435 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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This data was developed using ab300435, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD47 with ab300435 at 1/2000 dilution (0.244 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining on human tonsil. The section was incubated with ab300435 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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)
ab300436 は論文での使用が確認できていません。