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18350
Tyro/Axl/Mer Activation Sampler Kit
Primary Antibodies

Tyro/Axl/Mer Activation Sampler Kit #18350

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Western blot analysis of extracts from NCI-H1299 and SK-MEL-5 cells, untreated (-) or treated with GAS6 (400 ng/ml) and H2O2 (4 mM) for 10 min, using Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) (D6M4W) Rabbit mAb (upper) and Axl (C89E7) Rabbit mAb #8661 (lower 1), Mer (D21F11) XP® Rabbit mAb #4319 (lower 2), and Tyro3 (D38C6) Rabbit mAb #5585 (lower 3).

Western blot analysis of extracts from NCI-H1299 cells, untreated or Gas6-treated (100 ng/ml for 10 minutes), using Phospho-Axl (Tyr702) (D12B2) Rabbit mAb (upper) or Axl (C44G1) Rabbit mAb #4566 (lower).

Western blot analysis of extracts from various cell lines using Axl (C89E7) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western blot analysis of extracts from SK-MEL-5, Hep G2 and Jurkat cells using Mer (D21F11) XP® Rabbit mAb.

Western blot analysis of extracts from COS-7 cells, and tissues from human and mouse brain using Tyro3 (D38C6) Rabbit mAb.

Western blot analysis of extracts from various cell lines using GAS6 (D3A3G) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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.

Immunoprecipitation of Phospho-Axl (Tyr698) protein from NCI-H1299 cells treated with GAS6 (400 ng/ml) and H2O2 (4 mM) for 10 min. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, lane 3 is Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb, and lane 4 is Axl (C89E7) Rabbit mAb #8661. Western blot analysis was performed using Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb (upper) and Axl (C89E7) Rabbit mAb #8661 (lower). Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.

Immunohistochemical analysis of paraffin-embedded breast carcinoma using Axl (C89E7) Rabbit mAb. Note staining of infiltrating cells.

Confocal immunofluorescent analysis of SK-MEL-5 cells (left), Hep G2 cells (center) and MCF7 cells (right) using Mer (D21F11) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ® #4084 (fluorescent DNA dye).

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using GAS6 (D3A3G) Rabbit mAb.

Immunoprecipitation of Phospho-Mer (Tyr749) protein from SK-MEL-5 cells treated with GAS6 (400 ng/ml) and H2O2 (4 mM) for 10 min. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, lane 3 is Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb, and lane 4 is Mer(D21F11) XP® Rabbit mAb #4319. Western blot analysis was performed using Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb (upper) and Mer (D21F11) XP® Rabbit mAb #4319 (lower). Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.

Immunohistochemical analysis of paraffin-embedded metastatic lung carcinoma using Axl (C89E7) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded NCI-H226 cell pellet (left, positive) or HT29 cell pellet (right, negative) using GAS6 (D3A3G) Rabbit mAb.

Immunoprecipitation of Phospho-Tyro3 (Tyr681) protein from SK-MEL-5 cells treated with GAS6 (400 ng/ml) and H2O2 (4 mM) for 10 min. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, lane 3 is Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb, and lane 4 is Tyro 3 (D38C6) Rabbit mAb #5585. Western blot analysis was performed using Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb (upper) and Tyro3 (D38C6) Rabbit mAb #5585 (lower). Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.

Immunohistochemical analysis of paraffin-embedded cell pellets, NCI-H1299 (left) or Jurkat (right), using Axl (C89E7) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using GAS6 (D3A3G) Rabbit mAb.

Flow cytometric analysis of Jurkat cells (blue) and DU145 cells (green) using Axl (C89E7H4) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human serous carcinoma of the ovary using GAS6 (D3A3G) Rabbit mAb.

Confocal immunofluorescent analysis of DU 145 (left) and HCC827 (right) cells using Axl (C89E7) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Confocal immunofluorescent analysis of NCI-H226 (left, positive) and HT-29 (right, negative) using GAS6 (D3A3G) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

To Purchase # 18350T
Product # Size Price
18350T
1 Kit  (6 x 20 µl) N/A

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) (D6M4W) Rabbit mAb 44463 20 µl
  • WB
  • IP
H 110-130, 125-140, 180-200 Rabbit IgG
Phospho-Axl (Tyr702) (D12B2) Rabbit mAb 5724 20 µl
  • WB
H 138 Rabbit IgG
Axl (C89E7) Rabbit mAb 8661 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H Mk 138 Rabbit IgG
Mer (D21F11) XP® Rabbit mAb 4319 20 µl
  • WB
  • IP
  • IF
H 210 Rabbit IgG
Tyro3 (D38C6) Rabbit mAb 5585 20 µl
  • WB
  • IP
H M R Mk 110, 130 Rabbit IgG
GAS6 (D3A3G) Rabbit mAb 67202 20 µl
  • WB
  • IHC
  • IF
H 78 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Tyro/Axl/Mer Activation Sampler Kit provides an economical means of detecting the activation of TAM family members using phospho-specific and control antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) (D6M4W) Rabbit mAb detects immunoprecipitated or transfected levels of Axl/Mer/Tyro3 protein when phosphorylated at Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681). This antibody may cross-react with other phosphorylated tyrosine proteins. Phospho-Axl (Tyr702) (D12B2) Rabbit mAb detects endogenous levels of Axl only when phosphorylated at Tyr702. This antibody may also cross-react with other overexpressed, related tyrosine-phosphorylated tyrosine kinases. Axl (C89E7) Rabbit mAb, Mer (D21F11) XP® Rabbit mAb, Tyro3 (D38C6) Rabbit mAb, and GAS6 (D3A3G) Rabbit mAb detected endogenous levels of each of the corresponding target proteins.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to phospho-Tyr698/Tyr749 of human Axl/Mer, phospho-Tyr702 of human Axl, a recombinant fragment of human Axl, synthetic peptides corresponding to His925 of human Mer, Thr874 of human Tyro3, and recombinant protein specific to the amino terminus of human GAS6.

Background

Axl, Mer and Tyro3 are three members of the TAM family receptor tyrosine kinase that share a common NCAM (neural adhesion molecule)-related extracellular domain and a conserved intracellular tyrosine kinase domain. These receptors bind common homologous vitamin K dependent protein GAS6 and protein S to activate downstream signaling pathways (1). TAM family receptors are involved in the development of immune, nervous, vascular and reproductive systems, autoimmune disease, cancer drug resistance and tumor immunity response (2-5). Axl (Tyr698), Axl (Tyr702), Mer Tyr(749) and Tyro3 (Tyr681) are conserved autophosphorylation sites located in the activation loop of the respective tyrosine kinase domains. Phosphorylation at these sites is required for full kinase activation of each of the corresponding receptors (6,7).

  1. Rothlin, C.V. et al. (2015) Annu Rev Immunol 33, 355-91.
  2. Burstyn-Cohen, T. (2017) Int J Dev Biol 61, 215-24.
  3. Bellan, M. et al. (2016) Int J Mol Sci 17, pii: E1807.
  4. Schoumacher, M. and Burbridge, M. (2017) Curr Oncol Rep 19, 19.
  5. Akalu, Y.T. et al. (2017) Immunol Rev 276, 165-77.
  6. Ling, L. et al. (1996) J Biol Chem 271, 18355-62.
  7. Onken, J. et al. (2017) Oncotarget 8, 50403-14.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

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