Anti-Syk antibody [EP573Y] KO tested (ab40781)

Knockout Tested Rabbit Recombinant Monoclonal Syk antibody. Suitable for WB, IHC-P and reacts with Mouse, Human samples. Cited in 22 publications.

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Images

Western blot - Anti-Syk antibody [EP573Y] (AB40781), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Not recommended	Not recommended
Mouse	Tested	Expected	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/5000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000 - 1/5000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/5000	Notes For unpurified use at 1/100 - 1/250. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Species Mouse	Dilution info -	Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Species Rat	Dilution info -	Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Associated Products

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Target data

See full target information SYK

Function

Non-receptor tyrosine kinase which mediates signal transduction downstream of a variety of transmembrane receptors including classical immunoreceptors like the B-cell receptor (BCR). Regulates several biological processes including innate and adaptive immunity, cell adhesion, osteoclast maturation, platelet activation and vascular development (PubMed:12387735, PubMed:33782605). Assembles into signaling complexes with activated receptors at the plasma membrane via interaction between its SH2 domains and the receptor tyrosine-phosphorylated ITAM domains. The association with the receptor can also be indirect and mediated by adapter proteins containing ITAM or partial hemITAM domains. The phosphorylation of the ITAM domains is generally mediated by SRC subfamily kinases upon engagement of the receptor. More rarely signal transduction via SYK could be ITAM-independent. Direct downstream effectors phosphorylated by SYK include DEPTOR, VAV1, PLCG1, PI-3-kinase, LCP2 and BLNK (PubMed:12456653, PubMed:15388330, PubMed:34634301, PubMed:8657103). Initially identified as essential in B-cell receptor (BCR) signaling, it is necessary for the maturation of B-cells most probably at the pro-B to pre-B transition (PubMed:12456653). Activated upon BCR engagement, it phosphorylates and activates BLNK an adapter linking the activated BCR to downstream signaling adapters and effectors. It also phosphorylates and activates PLCG1 and the PKC signaling pathway. It also phosphorylates BTK and regulates its activity in B-cell antigen receptor (BCR)-coupled signaling. In addition to its function downstream of BCR also plays a role in T-cell receptor signaling. Plays also a crucial role in the innate immune response to fungal, bacterial and viral pathogens. It is for instance activated by the membrane lectin CLEC7A. Upon stimulation by fungal proteins, CLEC7A together with SYK activates immune cells inducing the production of ROS. Also activates the inflammasome and NF-kappa-B-mediated transcription of chemokines and cytokines in presence of pathogens. Regulates neutrophil degranulation and phagocytosis through activation of the MAPK signaling cascade (By similarity). Required for the stimulation of neutrophil phagocytosis by IL15 (PubMed:15123770). Also mediates the activation of dendritic cells by cell necrosis stimuli. Also involved in mast cells activation. Involved in interleukin-3/IL3-mediated signaling pathway in basophils (By similarity). Also functions downstream of receptors mediating cell adhesion (PubMed:12387735). Relays for instance, integrin-mediated neutrophils and macrophages activation and P-selectin receptor/SELPG-mediated recruitment of leukocytes to inflammatory loci. Also plays a role in non-immune processes. It is for instance involved in vascular development where it may regulate blood and lymphatic vascular separation. It is also required for osteoclast development and function. Functions in the activation of platelets by collagen, mediating PLCG2 phosphorylation and activation. May be coupled to the collagen receptor by the ITAM domain-containing FCER1G. Also activated by the membrane lectin CLEC1B that is required for activation of platelets by PDPN/podoplanin. Involved in platelet adhesion being activated by ITGB3 engaged by fibrinogen. Together with CEACAM20, enhances production of the cytokine CXCL8/IL-8 via the NFKB pathway and may thus have a role in the intestinal immune response (By similarity).

Alternative names

Tyrosine-protein kinase SYK, Spleen tyrosine kinase, p72-Syk, SYK

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal Syk antibody. Suitable for WB, IHC-P and reacts with Mouse, Human samples. Cited in 22 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EP573Y
Purification technique: Affinity purification Protein A
Specificity: The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

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

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.

Supplementary info

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

Activity summary

Spleen tyrosine kinase commonly known as Syk is a non-receptor tyrosine kinase with a mass of about 72 kDa. It serves as a critical signaling molecule in immune cells. Syk is expressed in various cell types such as B cells T cells monocytes and others involved in the immune response. It initiates signaling cascades by binding to immunoreceptor tyrosine-based activation motifs (ITAMs). Alternate forms include phosphorylated Syk often referred to as p-Syk which activates various downstream signaling pathways.

Biological function summary

Syk plays a role in transmitting signals from the cell surface to the interior of the cell facilitating various cellular activities. It is a part of signal transduction complexes and functions in the regulation of immune cell activation differentiation and survival. It affects processes like Fc receptor signaling in macrophages and neutrophils impacting immune responses and inflammation. The phospho-Syk form influences the strength and duration of signaling affecting various cellular functions.

Pathways

Syk is significant in the B-cell receptor (BCR) signaling pathway and Fc receptor signaling pathway. Within these pathways Syk interacts with proteins like Lyn and Fyn which are other kinases involved in BCR signal transduction. Syk phosphorylation noted as p-Syk plays a part in activating downstream molecules such as phospholipase C gamma (PLCγ) to mediate cellular responses to external stimuli. The critical role of Syk in these pathways influences both adaptive and innate immune responses.

Associated diseases and disorders

Syk correlates with conditions like rheumatoid arthritis and certain B-cell lymphomas. In rheumatoid arthritis aberrant Syk signaling contributes to the inflammation and joint destruction characteristic of the disease. Syk's role in B-cell antigen receptor signaling links it to B-cell lymphomas where dysregulation in signaling pathways leads to uncontrolled proliferation of malignant B cells. The involvement of Syk in both disease contexts suggests its potential as a therapeutic target along with pathways related to proteins like BLNK and PLCγ in these disorders.

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10 product images

Western blot - Anti-Syk antibody [EP573Y] (ab40781)
False colour image of Western blot: Anti-Syk antibody [EP573Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab40781 was shown to bind specifically to Syk. A band was observed at 70/72 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SYK knockout cell line Human SYK knockout HEK-293T cell line ab282649 (knockout cell lysate Human SYK knockout HEK-293T cell lysate ab283048). To generate this image, wild-type and SYK knockout HEK-293T 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Syk antibody [EP573Y] (ab40781) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: SYK knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human SYK knockout HEK-293T cell line (Human SYK knockout HEK-293T cell line ab282649)
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 70 kDa
Western blot - Anti-Syk antibody [EP573Y] (ab40781)
Lane 1: Wild-type HAP1 cell lysate (40 μg)
Lane 2: SYK knockout HAP1 cell lysate (40 μg)
Lane 3: K562 cell lysate (40 μg)
Lane 4: Raji cell lysate (40 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab40781 observed at 75 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab40781 was shown to specifically react with Syk when Syk knockout samples were used. Wild-type and Syk knockout samples were subjected to SDS-PAGE. ab40781 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted at 1/5000 and 1/10000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Syk antibody [EP573Y] (ab40781)
Predicted band size: 72 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syk antibody [EP573Y] (ab40781)
Immunohistochemistry of paraffin embedded Human spleen tissue section labelling Syk with ab40781 at 1:5000 dilution (0.44 μg/ml). Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). A ready to use ImmunoHistoProbe one step HRP Polymer (ready to use) was used as a secondary antibody at 1:0 dilution. PBS instead of primary antibody was used for negative control. Hematoxylin was used as a counterstain.
Western blot - Anti-Syk antibody [EP573Y] (ab40781)
Blocking/Diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Syk antibody [EP573Y] (ab40781) at 1/1000 dilution
Lane 1: Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 15 µg
Lane 2: Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysates at 15 µg
Lane 3: WEHI-231 (Mouse B cell lymphoma B lymphocyte ) whole cell lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 72 kDa
Observed band size: 72 kDa
Western blot - Anti-Syk antibody [EP573Y] (ab40781)
Blocking/Diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Syk antibody [EP573Y] (ab40781) at 1/1000 dilution
All lanes: Human bone marrow lysates at 15 µg
Secondary
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 72 kDa
Observed band size: 72 kDa
Western blot - Anti-Syk antibody [EP573Y] (ab40781)
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Syk antibody [EP573Y] (ab40781) at 1/5000 dilution
All lanes: Mouse bone marrow lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 72 kDa
Exposure time: 3min
Western blot - Anti-Syk antibody [EP573Y] (ab40781)
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Syk antibody [EP573Y] (ab40781) at 1/2000 dilution
All lanes: Rat bone marrow lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 72 kDa
Exposure time: 50s
Western blot - Anti-Syk antibody [EP573Y] (ab40781)
Western blot: Anti-SYK antibody [EP573Y] (ab40781) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab40781 was shown to bind specifically to SYK. A band was observed at 72 kDa in wild-type THP-1 cell lysates with no signal observed at this size in SYK knockout cell line Human SYK knockout THP-1 cell line ab288700 (knockout cell lysate ab289593). To generate this image, wild-type and SYK knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Syk antibody [EP573Y] (ab40781) at 1/1000 dilution
Lanes 1 - 4: Western blot at 20 µg
Lane 2: Western blot - Human SYK knockout THP-1 cell line (Human SYK knockout THP-1 cell line ab288700)
Secondary
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 72 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Syk antibody [EP573Y] (ab40781)
ab40781 was shown to react with SYK in wild-type THP-1 cells in immunocytochemistry with loss of signal observed in SYK knockout cell line Human SYK knockout THP-1 cell line ab288700. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab40781 at dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
Western blot - Anti-Syk antibody [EP573Y] (ab40781)
ab40781 was shown to react with SYK in wild-type THP-1 cells in Western blot with loss of signal observed in SYK knockout cell line Human SYK knockout THP-1 cell line ab288700. Wild-type THP-1 and SYK knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab40781 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-Syk antibody [EP573Y] (ab40781) at 1/1000 dilution
Lane 1: Wild-type THP-1 lysate at 20 µg
Lane 2: Western blot - Human SYK knockout THP-1 cell line (Human SYK knockout THP-1 cell line ab288700) at 20 µg
Observed band size: 72 kDa