Anti-Fyn 抗体 [EPR19636] - BSA and Azide free (ab246333)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19636] to Fyn - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IHC-P, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Fyn antibody [EPR19636] - BSA and Azide free
Fyn 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR19636] to Fyn - BSA and Azide free -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), IHC-P, IP, WBmore 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: Human brain lysate; Ramos, HEK-293, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; P0 mouse brain lysate; P0 rat brain lysate; Rat spleen lysate. IHC-P: Human tonsil and cerebral cortex tissues; Mouse spleen tissue; Rat testis tissue. Flow Cyt (intra): RAW 264.7 cells. IP: P0 mouse brain lysate.
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特記事項
ab246333 is the carrier-free version of ab184276.
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
EPR19636 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- Anti-Fyn antibody [EPR19636] (ab184276)
- Alexa Fluor® 488 Anti-Fyn antibody [EPR19636] (ab309857)
- Alexa Fluor® 647 Anti-Fyn antibody [EPR19636] (ab310225)
- Alexa Fluor® 594 Anti-Fyn antibody [EPR19636] (ab310673)
- Alexa Fluor® 555 Anti-Fyn antibody [EPR19636] (ab312203)
- Alexa Fluor® 568 Anti-Fyn antibody [EPR19636] (ab312692)
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Compatible Secondaries
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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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Recombinant Protein
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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ab246333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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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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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
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特記事項 |
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Flow Cyt (Intra)
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. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa). |
ターゲット情報
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機能
Tyrosine-protein kinase implicated in the control of cell growth. Plays a role in the regulation of intracellular calcium levels, with isoform 2 showing the greater ability to mobilize cytoplasmic calcium in comparison to isoform 1. Required in brain development and mature brain function with important roles in the regulation of axon growth, axon guidance, and neurite extension. Blocks axon outgrowth and attraction induced by NTN1 by phosphorylating its receptor DDC. Phosphorylates RUNX3. -
組織特異性
Isoform 1 is highly expressed in the brain. Isoform 2 is expressed in cells of hemopoietic lineages, especially T lymphocytes. -
配列類似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
Contains 1 protein kinase domain.
Contains 1 SH2 domain.
Contains 1 SH3 domain. -
細胞内局在
Cell membrane. Present and active in lipid rafts. Present in cell body and along the process of mature and developing oligodendroyctes. - Information by UniProt
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参照データベース
- Entrez Gene: 2534 Human
- Entrez Gene: 14360 Mouse
- Entrez Gene: 25150 Rat
- Omim: 137025 Human
- SwissProt: P06241 Human
- SwissProt: P39688 Mouse
- SwissProt: Q62844 Rat
- Unigene: 390567 Human
see all -
製品の状態
This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), YES1 (P07947), SRC (P12931), and LYN (P07948). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request. -
別名
- C syn protooncogene antibody
- Fyn antibody
- FYN oncogene related to SRC FGR YES antibody
see all
画像
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All lanes : Anti-Fyn antibody [EPR19636] (ab184276) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 cell lysate
Lane 2 : FYN knockout HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Fyn antibody [EPR19636] 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, ab184276 was shown to bind specifically to Fyn. A band was observed at 60 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in FYN knockout cell line ab269630 (knockout cell lysate ab272440). To generate this image, wild-type and FYN knockout HEK-293 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-Fyn antibody [EPR19636] (ab184276) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : FYN knockout HEK-293T cell lysate
Lane 3 : Human brain tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 159 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 (ab184276).
Lanes 1-3: Merged signal (red and green). Green - ab184276 observed at 60 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab184276 Anti-Fyn antibody [EPR19636] was shown to specifically react with Fyn in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266133 (knockout cell lysate ab257071) was used. Wild-type and Fyn knockout samples were subjected to SDS-PAGE. ab184276 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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 10000 dilution for 1 hour at room temperature before imaging.
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Fyn was immunoprecipitated from 0.35 mg of P0 mouse brain lysate with ab184276 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab184276 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: P0 mouse brain lysate 10 µg (Input).
Lane 2: ab184276 IP in P0 mouse brain lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184276 in P0 mouse brain lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184276).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line labeling Fyn with ab184276 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184276).
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on T cells of human tonsil (PMID: 10523617). 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184276).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human cerebral cortex (PMID: 7544314). 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184276).Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on T cells of mouse spleen (PMID: 7544314, PMID:10523617). 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184276).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on rat testis (PMID: 7544314). 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184276).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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)
ab246333 は論文での使用が確認できていません。