AB300398

Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) 抗体 [EPR26232-39]

Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39]

BOND RX™ Validated
20ul selling size
RabMAb
Recombinant
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Rabbit Recombinant Monoclonal ZAP70 phospho Y319 antibody. Suitable for WB, Dot, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Synthetic peptide - Human, Synthetic peptide - Mouse samples.

別名を表示する

SRK, ZAP70, Tyrosine-protein kinase ZAP-70, 70 kDa zeta-chain associated protein, Syk-related tyrosine kinase, Tyrosine-protein kinase SYK, Spleen tyrosine kinase, p72-Syk, SYK

9 Images

Immunocytochemistry/ Immunofluorescence - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat (human T cell leukemia T lymphocyte) cells labelling Syk (phospho Y352) +ZAP70 (phospho Y319) with ab300398 at 1/50 (9.44 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing increased membranous staining in Jurkat cells treated with pervanadate (50 mM) for 5 min is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

Flow Cytometry (Intracellular) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) treated with 1mM pervanadate for 30 minutes (Red)/ Untreated control (Green) cells labelling Syk (phospho Y352) +ZAP70 (phospho Y319) with ab300398 at 1/500 dilution (0.1ug) (Red) and Green (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Immunohistochemical analysis of paraffin-embedded Panel A, Mouse splee tissue labeling Syk (phospho Y352) +ZAP70 (phospho Y319) with ab300398 at 1/500 (0.944 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse spleen without alkaline phosphatase treatment (Panel A); no staining on mouse spleen with alkaline phosphatase treatment (Panel B). The section was incubated with ab300398 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized EL4 (mouse lymphoma t lymphocyte) treated with 11mM H2O2 for 10 minutes then 10mM pervanadate for 30 minutes (Red)/ Untreated control (Green) cells labelling Syk (phospho Y352) +ZAP70 (phospho Y319) with ab300398 at 1/50 dilution (1ug) (Red) and Green (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized EL4 (mouse lymphoma T lymphocyte) cells labelling Syk (phospho Y352) +ZAP70 (phospho Y319) with ab300398 at 1/50 (9.44 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing increased membranous staining in EL4 cells treated with H2O2 (11 mM) for 10 min and then pervanadate (10 mM) for 30 min is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Immunohistochemical analysis of paraffin-embedded Panel A, Mouse pancr tissue labeling Syk (phospho Y352) +ZAP70 (phospho Y319) with ab300398 at 1/500 (0.944 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse pancreatic cancer without alkaline phosphatase treatment (Panel A); no staining on mouse pancreatic cancer with alkaline phosphatase treatment (Panel B). The section was incubated with ab300398 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Immunohistochemical analysis of paraffin-embedded Panel A, Mouse lung tissue labeling Syk (phospho Y352) +ZAP70 (phospho Y319) with ab300398 at 1/500 (0.944 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse lung without alkaline phosphatase treatment (Panel A); no staining on mouse lung with alkaline phosphatase treatment (Panel B). The section was incubated with ab300398 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Supplier Data

Western blot - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Blocking / Diluting buffer and concentration : 5% NFDM/TBST

Exposure time : Lanes 1-4 : 6 seconds; Lanes 5-6 : 147 seconds.

All lanes:

Western blot - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (ab300398) at 1/1000 dilution

Lane 1:

Untreated Jurkat whole cell lysate at 20 µg

Lane 2:

Jurkat treated with 50mM pervanadate for 5 minutes whole cell lysate at 20 µg

Lane 3:

ntreated starved 24 hours Ramos whole cell lysate at 20 µg

Lane 4:

Ramos starved 24 hours, then treated with 10mM pervanadate for 30 minuteswhole cell lysate at 20 µg

Lane 5:

Untreated EL4 whole cell lysate at 20 µg

Lane 6:

EL4 treated with 11mM H2O2 for 10 minutes, then 10mM pervanadate for 30 minutes whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 70 kDa,72 kDa

false

Supplier Data

Dot Blot - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Blocking/diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Dot Blot - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (ab300398) at 1/1000 dilution

Lane 1:

Syk (phospho Y352) peptide a

Lane 2:

Syk (phospho Y352) peptide b

Lane 3:

Syk non-phospho peptide

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>)

false

Exposure time: 180s

Key facts

宿主種

Rabbit

クローン性

Monoclonal

クローン番号

EPR26232-39

アイソタイプ

IgG

キャリアフリー

交差種

Human, Mouse

アプリケーション

Flow Cyt (Intra), ICC/IF, WB, IHC-P, Dot

applications

免疫原

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "1/500", "IHCP-species-notes": "Not suitable for Human Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/50", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/50", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Mouse": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/500", "IHCP-species-notes": "Not suitable for Human Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/50", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/50", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Rat": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "1/500", "IHCP-species-notes": "Not suitable for Human Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Synthetic peptide - Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Synthetic peptide - Mouse": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" } } }

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製品の詳細

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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出荷温度及び保存条件

製品の状態

Liquid

精製方法

Affinity purification Protein A

バッファー組成

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

出荷温度

Conditional Ambient

短期保存期間

1-2 weeks

短期保存温度

+4°C

長期保存温度

-20°C

分注に関する情報

Upon delivery aliquot

保管に関する情報

Do Not Freeze

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

Syk (spleen tyrosine kinase) and ZAP70 (zeta-chain-associated protein kinase 70) are non-receptor tyrosine kinases. Syk has a molecular mass of approximately 72 kDa while ZAP70 is around 70 kDa. They are expressed in hematopoietic tissues. Syk is found in B-cells and other myeloid lineages and ZAP70 is mainly located in T-cells and natural killer (NK) cells. These kinases play essential roles in signal transduction by phosphorylating proteins on tyrosine residues transmitting activation signals from receptors on the cell surface to the interior of the cell.

Biological function summary

Syk and ZAP70 play vital roles in the adaptive immune response. Syk is an essential mediator of B-cell receptor (BCR) signaling and assembles into a signaling complex with other proteins. ZAP70 functions similarly in T-cells becoming activated upon the engagement of the T-cell receptor (TCR). Both kinases help recruit and activate adaptor proteins and downstream signaling molecules that drive cellular responses including proliferation differentiation and survival.

Pathways

Both kinases are integral to the B-cell and T-cell receptor signaling pathways. Syk activates several pathways including the MAP kinase and NF-kB pathways through its interaction with proteins like BLNK and PLCγ2. ZAP70 initiates the TCR signaling cascade interfacing with LAT and SLP-76 to activate the same pathways. These pathways trigger transcriptional programs necessary for immune cell functions and development.

Syk and ZAP70 have connections to various immune-related conditions. Abnormal Syk activity relates to autoimmune diseases like rheumatoid arthritis where B-cell signaling plays a role. Syk interacts with proteins such as ITAM-containing immunoreceptors in these disorders. ZAP70 mutations are linked to immunodeficiency syndromes leading to dysfunctional T-cell signaling. These kinases make viable targets for therapeutic intervention bringing hope for better management of such immune diseases.

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製品プロトコール

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

Tyrosine kinase that plays an essential role in regulation of the adaptive immune response. Regulates motility, adhesion and cytokine expression of mature T-cells, as well as thymocyte development. Also contributes to the development and activation of primary B-lymphocytes. When antigen presenting cells (APC) activate T-cell receptor (TCR), a serie of phosphorylations lead to the recruitment of ZAP70 to the phosphorylated TCR components CD3E and CD247/CD3Z through ITAM motif at the plasma membrane (PubMed : 7509083). This recruitment serves to localization to the stimulated TCR and to relieve its autoinhibited conformation. Release of ZAP70 active conformation is further stabilized by phosphorylation mediated by LCK. Subsequently, ZAP70 phosphorylates at least 2 essential adapter proteins : LAT and LCP2. In turn, a large number of signaling molecules are recruited and ultimately lead to lymphokine production, T-cell proliferation and differentiation. Furthermore, ZAP70 controls cytoskeleton modifications, adhesion and mobility of T-lymphocytes, thus ensuring correct delivery of effectors to the APC. ZAP70 is also required for TCR-CD247/CD3Z internalization and degradation through interaction with the E3 ubiquitin-protein ligase CBL and adapter proteins SLA and SLA2. Thus, ZAP70 regulates both T-cell activation switch on and switch off by modulating TCR expression at the T-cell surface. During thymocyte development, ZAP70 promotes survival and cell-cycle progression of developing thymocytes before positive selection (when cells are still CD4/CD8 double negative). Additionally, ZAP70-dependent signaling pathway may also contribute to primary B-cells formation and activation through B-cell receptor (BCR).

See full target information ZAP70 pY319

追加のターゲット

SYK phospho Y352

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Abcam product promise

当社は、高品質な試薬を通じてお客様の研究を力強くサポートすることをお約束いたします。ご使用いただく各段階で、常にお客様をサポートできる体制を整えております。万が一、製品が期待通りに機能しない場合は、「Abcam Product Promise」による当社保証制度に基づき、安心してご利用いただけます。

保証に関する詳細については利用規約をご確認ください。

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) 抗体 [EPR26232-39]

Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39]

Immunocytochemistry/ Immunofluorescence - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Flow Cytometry (Intracellular) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Flow Cytometry (Intracellular) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Immunocytochemistry/ Immunofluorescence - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Western blot - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

Dot Blot - Anti-Syk (phospho Y352) +ZAP70 (phospho Y319) antibody [EPR26232-39] (AB300398)

関連する標識済み抗体及び組成の異なる製品 (1)

Key facts

宿主種

クローン性

クローン番号

アイソタイプ

キャリアフリー

交差種

アプリケーション

免疫原

組合せ製品

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

製品の詳細

出荷温度及び保存条件

製品の状態

精製方法

バッファー組成

出荷温度

短期保存期間

短期保存温度

長期保存温度

分注に関する情報

保管に関する情報

補足情報

Biological function summary

Pathways

製品プロトコール

ターゲットの情報

追加のターゲット

Abcam product promise