Revision 6

#18350Store at -20C

1 Kit

(6 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
Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) (D6M4W) Rabbit mAb 44463 20 µl 110-130, 125-140, 180-200 kDa Rabbit IgG
Phospho-Axl (Tyr702) (D12B2) Rabbit mAb 5724 20 µl 138 kDa Rabbit IgG
Axl (C89E7) Rabbit mAb 8661 20 µl 138 kDa Rabbit IgG
Mer (D21F11) XP® Rabbit mAb 4319 20 µl 210 kDa Rabbit IgG
Tyro3 (D38C6) Rabbit mAb 5585 20 µl 110, 130 kDa Rabbit IgG
GAS6 (D3A3G) Rabbit mAb 67202 20 µl 78 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 Tyro/Axl/Mer Activation Antibody 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.

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

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.

Background References

    Trademarks and Patents

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

    使用に関する制限

    法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    Revision 6
    #18350

    Tyro/Axl/Mer Activation Antibody Sampler Kit

    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 1 Expand Image
    Flow cytometric analysis of MCF7 cells (blue) and HepG2 cells (green) using Mer (D21F11) XP® Rabbit mAb (solid lines) or a concetration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 2 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using Axl (C89E7) Rabbit mAb #8661. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from SK-MEL-5, Hep G2 and Jurkat cells using Mer (D21F11) XP® Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 4 Expand Image
    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).
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from COS-7 cells, and tissues from human and mouse brain using Tyro3 (D38C6) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 6 Expand Image
    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).
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various cell lines using GAS6 (D3A3G) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 8 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.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of HeLa Cell Extracts, untreated (-) or Axl knock-out (+) using Axl (C89E7) Rabbit mAb, #8661 (upper) or GAPDH (D16H11) XP® Rabbit mAb, #5174 (lower).
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 10 Expand Image
    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).
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 11 Expand Image
    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.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using GAS6 (D3A3G) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from various cell lines using Axl (C89E7) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 14 Expand Image
    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.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 15 Expand Image
    Immunohistochemical analysis of paraffin-embedded NCI-H226 cell pellet (left, positive) or HT29 cell pellet (right, negative) using GAS6 (D3A3G) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 16 Expand Image
    Immunohistochemical analysis of paraffin-embedded breast carcinoma using Axl (C89E7) Rabbit mAb. Note staining of infiltrating cells.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 17 Expand Image
    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.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using GAS6 (D3A3G) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using Axl (C89E7) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded human serous carcinoma of the ovary using GAS6 (D3A3G) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using Axl (C89E7) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 22 Expand Image
    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).
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using Axl (C89E7) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded metastatic lung carcinoma using Axl (C89E7) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded cell pellets, NCI-H1299 (left) or Jurkat (right), using Axl (C89E7) Rabbit mAb.
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 26 Expand Image
    Confocal immunofluorescent analysis of DU 145 (left) and HCC827 (right) cells using Axl (C89E7) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Tyro/Axl/Mer Activation Antibody Sampler Kit: Image 27 Expand Image
    Flow cytometric analysis of fixed and permeabilized Jurkat cells (blue, negative) and DU145 cells (green, positive) using Axl (C89E7H4) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.