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97999
Phospho-Jak Family Antibody Sampler Kit

Phospho-Jak Family Antibody Sampler Kit #97999

Western Blotting Image 1

Western blot analysis of extracts from ACHN, SR, CTLL-2 and PC12 cell lines using Jak1 (6G4) Rabbit mAb.

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Western Blotting Image 2

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).

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Western Blotting Image 3

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.

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Western Blotting Image 4

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).

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Western Blotting Image 5

Western blot analysis of extracts from various cell lines using Jak3 (D1H3) Rabbit mAb.

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Western Blotting Image 6

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).

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Western Blotting Image 7

Western blot analysis of extracts from various cell lines using Tyk2 (D4I5T) Rabbit mAb.

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Western Blotting Image 8

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).

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Western Blotting Image 9

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.

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IHC-P (paraffin) Image 10

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).

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Western Blotting Image 11

Western blot analysis of extracts from K-562, THP-1, TF-1 and BaF3 cell lines using Jak2 (D2E12) XP® Rabbit mAb.

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Western Blotting Image 12

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).

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Western Blotting Image 13

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.

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IHC-P (paraffin) Image 14

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Jak1 (6G4) Rabbit mAb.

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IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Jak2 (D2E12) XP® Rabbit mAb.

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Western Blotting Image 16

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).

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Western Blotting Image 17

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.

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IHC-P (paraffin) Image 18

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Jak1 (6G4) Rabbit mAb.

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IHC-P (paraffin) Image 19

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).

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Flow Cytometry Image 20

Flow cytometric analysis of K-562 cells using Jak2 (D2E12) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IF-IC Image 21

Confocal immunofluorescent analysis of K-562 cells using Jak2 (D2E12) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Jak1 (6G4) Rabbit mAb 3344 20 µl
  • WB
  • IHC
H M R 130 Rabbit IgG
Phospho-Jak1(Tyr1034/1035) (D7N4Z) Rabbit mAb 74129 20 µl
  • WB
H M R 130 Rabbit IgG
Jak2 (D2E12) XP® Rabbit mAb 3230 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R 125 Rabbit IgG
Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb 8082 20 µl
  • WB
H M 125 Rabbit IgG
Jak3 (D1H3) Rabbit mAb 8827 20 µl
  • WB
  • IP
H 115 Rabbit IgG
Phospho-Jak3 (Tyr980/981) (D44E3) Rabbit mAb 5031 20 µl
  • WB
H M 115 Rabbit IgG
Tyk2 (D4I5T) Rabbit mAb 14193 20 µl
  • WB
  • IP
H Mk 134 Rabbit IgG
Phospho-Tyk2 (Tyr1054/1055) (D7T8A) Rabbit mAb 68790 20 µl
  • WB
H 134 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

Each antibody in the Phospho-Jak Family Antibody Sampler kit detects endogenous levels of its target protein. Phospho-Jak1 (Tyr1034/1035) (D7N4Z) Rabbit mAb can detect Jak1 when dually or singly phosphorylated at Tyr1034. This site has historically been referenced as Tyr1022 and Ty1023. Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb can detect Jak2 when phosphorylated at Tyr1008 or when dually phosphorylated at Tyr1007 and Tyr1008. Phospho-Jak3 (Tyr980/981) (D44E3) Rabbit mAb can also detect Jak3 when dually phosphorylated or when phosphorylated at each of the single phosphorylation sites. Phospho-Tyk2 (Tyr1054/1055) (D7T8A) Rabbit mAb detects Tyk2 when phosphorylated at both Tyr1054 and Tyr1055. Cross-reactivity was not observed with other family members.

Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to Ile800 of Jak1, Pro841 of Jak2, and the carboxy terminus of Jak3 and Tyk2. Phosphorylation-specific monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to Tyr1034/1035 of Jak1, Tyr1008 of Jak2, Tyr980/981 of Jak3, and Tyr1054/1055 of Tyk2.

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.
Entrez-Gene Id
3716 , 3717 , 3718 , 7297
Swiss-Prot Acc.
P23458 , O60674 , P52333 , P29597
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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