Revision 1

#97999Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Jak1 (6G4) Rabbit mAb 3344 20 µl 130 kDa Rabbit IgG
Phospho-Jak1(Tyr1034/1035) (D7N4Z) Rabbit mAb 74129 20 µl 130 kDa Rabbit IgG
Jak2 (D2E12) XP® Rabbit mAb 3230 20 µl 125 kDa Rabbit IgG
Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb 8082 20 µl 125 kDa Rabbit IgG
Jak3 (D1H3) Rabbit mAb 8827 20 µl 115 kDa Rabbit IgG
Phospho-Jak3 (Tyr980/981) (D44E3) Rabbit mAb 5031 20 µl 115 kDa Rabbit IgG
Tyk2 (D4I5T) Rabbit mAb 14193 20 µl 134 kDa Rabbit IgG
Phospho-Tyk2 (Tyr1054/1055) (D7T8A) Rabbit mAb 68790 20 µl 134 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Phospho-Jak Family Antibody Sampler Kit provides an economical means of detecting the activation of Jak family members using phospho-specific and control antibodies. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Members of the Janus family of tyrosine kinases (Jak1, Jak2, Jak3, and Tyk2) are activated by ligands binding to a number of associated cytokine receptors (1). Upon cytokine receptor activation, Jak proteins become autophosphorylated and phosphorylate their associated receptors to provide multiple binding sites for signaling proteins. These associated signaling proteins, such as Stats (2), Shc (3), insulin receptor substrates (4), and focal adhesion kinase (FAK) (5), typically contain SH2 or other phosphotyrosine-binding domains.
Activation of Jak kinases upon cytokine receptor binding is associated with tyrosine phosphorylation within their activation loops, including Tyr1034/1035 of Jak1, Tyr1007/1008 of Jak2, Tyr980/981 of Jak3, and Tyr1054/1055 of Tyk2. Many studies have indicated that various cytokine receptors have clear preferences that utilize distinct Jak family members. Aberrant regulation of Jak signaling is associated with a number of diseases, including myeloproliferative neoplasms, leukemia, and inflammatory disease (6).

  1. Leonard, W.J. and O'Shea, J.J. (1998) Annu Rev Immunol 16, 293-322.
  2. Darnell, J.E. (1997) Science 277, 1630-5.
  3. VanderKuur, J. et al. (1995) J Biol Chem 270, 7587-93.
  4. Argetsinger, L.S. et al. (1995) J Biol Chem 270, 14685-92.
  5. Zhu, T. et al. (1998) J Biol Chem 273, 10682-9.
  6. Babon, J.J. et al. (2014) Biochem J 462, 1-13.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #97999

    Phospho-Jak Family Antibody Sampler Kit

    Phospho-Jak Family Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts from various cell lines using Tyk2 (D4I5T) Rabbit mAb.
    Phospho-Jak Family Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from various cell lines using Jak2 (D2E12) XP® Rabbit mAb #3230 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Phospho-Jak Family Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from ACHN, SR, CTLL-2 and PC12 cell lines using Jak1 (6G4) Rabbit mAb.
    Phospho-Jak Family Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from NK-92 cells, untreated or treated with Human Interleukin-2 (hIL-2) #8907 (10 ng/ml, 15 minutes), using Phospho-Jak3 (Tyr980/981) (D44E3) Rabbit mAb (upper) or Jak3 Antibody #3775 (lower).
    Phospho-Jak Family Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from serum-starved U266 and Jurkat cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (50 ng/ml, 15 min; +) using Phospho-Tyk2 (Tyr1054/1055) (D7T8A) Rabbit mAb (upper) or total Tyk2 (D4I5T) Rabbit mAb #14193 (lower).
    Phospho-Jak Family Antibody Sampler Kit: Image 6 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    Phospho-Jak Family Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various cell lines, serum-starved overnight (-) followed by treatment with Human Interferon-α1 #8927 (hIFN-α1, 10 ng/ml, 15 min; +) or Human Interleukin-4 #8919 (hIL-4, 10 ng/ml, 10 min; +) using Phospho-Jak1 (Tyr1034/1035) (D7N4Z) Rabbit mAb (upper) or Jak1 (6G4) Rabbit mAb (lower).
    Phospho-Jak Family Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from BaF3 cells, untreated or treated with Mouse Interleukin-3 (mIL-3) #8923 (10 ng/ml, 5 min), using Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb (upper) or total Jak2 (D2E12) XP® Rabbit mAb #3230 (lower).
    Phospho-Jak Family Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell lines using Jak3 (D1H3) Rabbit mAb.
    Phospho-Jak Family Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human Tyk2 (hTyk2; +), using Tyk2 (D4I5T) Rabbit mAb.
    Phospho-Jak Family Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from K-562 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or Jak2 siRNA (+), using Jak2 (D2E12) XP® Rabbit mAb #3230 and α-Tubulin (11H10) Rabbit mAb #2125. The Jak2 (D2E12) XP® Rabbit mAb confirms silencing of Jak2 expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Jak2 siRNA.
    Phospho-Jak Family Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Jak1 (6G4) Rabbit mAb #3344, in the presence of control peptide (left) or Jak1 Blocking Peptide (right).
    Phospho-Jak Family Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from SET-2 cells, untreated or treated with the Jak2 inhibitor FLLL31 (10 μM, 4 hr), using Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb (upper) or total Jak2 (D2E12) XP® Rabbit mAb #3230 (lower).
    Phospho-Jak Family Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from MCF7 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Tyk2 siRNA #14254 (+), using Tyk2 (D4I5T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The Tyk2 (D4I5T) Rabbit mAb confirms silencing of Tyk2 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
    Phospho-Jak Family Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of extracts from K-562, THP-1, TF-1 and BaF3 cell lines using Jak2 (D2E12) XP® Rabbit mAb.
    Phospho-Jak Family Antibody Sampler Kit: Image 16 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Jak1 (6G4) Rabbit mAb.
    Phospho-Jak Family Antibody Sampler Kit: Image 17 Expand Image
    Western blot analysis of extracts from LNCaP cells, untreated or treated with growth hormone (GH) (100 ng/ml, 5 min), using Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb (upper) or total Jak2 (D2E12) XP® Rabbit mAb #3230 (lower).
    Phospho-Jak Family Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Jak2 (D2E12) XP® Rabbit mAb.
    Phospho-Jak Family Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Jak1 (6G4) Rabbit mAb.
    Phospho-Jak Family Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Jak2 (D2E12) XP® Rabbit mA in the presence of control peptide (left) or Jak2 Blocking Peptide #1039 (right).