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Western blot
ImmunohistochemistryAntibody data
- Antibody Data
- Antigen structure
- References [3]
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- Validations
- Western blot [1]
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- Product number
- 14-9752-82 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ITK Monoclonal Antibody (2F12), eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The monoclonal antibody 2F12 recognizes human, mouse, and rat ITK. ITK (also known as Interleukin-2 inducible T-cell kinase, T-cell-specific kinase, Tyrosine-protein kinase Lyk, Emt, and Tsk) is a member of the Tec family of kinases that include Tec, Btk, Bmx, and Txk. Tec family kinases play an indispensable role in activation of phospholipase C and activation of calcium-dependent signaling cascades following antigen receptor triggering. ITK is highly expressed by T cells and is required for optimal TCR-induced PLCgamma1 activation. Mice congenitally lacking ITK show profound defects in T cell development and signaling. Defects in ITK are the cause of lymphoproliferative syndrome EBV-associated autosomal type 1 (LPSA1). ITK also plays a role in the development of various T cell mediated inflammatory disorders such as atopic dermatitis, psoriasis, and allergic asthma. Applications Reported: This 2F12 antibody has been reported for use in western blotting, immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, and microscopy. Applications Tested: This 2F12 antibody has been tested by immunohistochemistry of formalin-fixed paraffin embedded human tissue using high pH antigen retrieval and can be used at less than or equal to 1 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 2F12
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- 4° C
Submitted references Regulation of T-cell responses and disease by tec kinase Itk.
Girls homozygous for an IL-2-inducible T cell kinase mutation that leads to protein deficiency develop fatal EBV-associated lymphoproliferation.
The Tec family tyrosine kinases Itk and Rlk regulate the development of conventional CD8+ T cells.
August A, Ragin MJ
International reviews of immunology 2012 Apr;31(2):155-65
International reviews of immunology 2012 Apr;31(2):155-65
Girls homozygous for an IL-2-inducible T cell kinase mutation that leads to protein deficiency develop fatal EBV-associated lymphoproliferation.
Huck K, Feyen O, Niehues T, Rüschendorf F, Hübner N, Laws HJ, Telieps T, Knapp S, Wacker HH, Meindl A, Jumaa H, Borkhardt A
The Journal of clinical investigation 2009 May;119(5):1350-8
The Journal of clinical investigation 2009 May;119(5):1350-8
The Tec family tyrosine kinases Itk and Rlk regulate the development of conventional CD8+ T cells.
Atherly LO, Lucas JA, Felices M, Yin CC, Reiner SL, Berg LJ
Immunity 2006 Jul;25(1):79-91
Immunity 2006 Jul;25(1):79-91
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using Anti-ITK Monoclonal Antibody (2F12), eBioscience™ (Product # 14-9752-82) and a 71 kDa band corresponding to Tyrosine-protein kinase ITK/TSK was detected in the T-cell (Jurkat and MOLT-4) and natural killer cell line (NK-92) but not in B-cell lines (Raji and Ramos) and THP-1 monocyte. Whole cell extracts (40 µg lysate) of Jurkat (Lane 1), MOLT-4 (Lane 2), Raji (Lane 3), Ramos (Lane 4), THP-1 (Lane 5), NK-92 (Lane 6), Mouse Heart (Lane 7), Mouse Skeletal Muscle (Lane 8) and Mouse Thymus (Lane 9) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). The band detected at ~25 kDa corresponds to the free circulating IgG detected in mouse tissues.
