Revision 4

#3860Store at -20C

1 Kit

(5 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-PLCγ2 (Tyr1217) Antibody 3871 20 µl 150 kDa Rabbit 
Phospho-PLCγ2 (Tyr759) (E9E9Y) Rabbit mAb 50535 20 µl 150 kDa Rabbit IgG
PLCγ2 (E5U4T) Rabbit mAb 55512 20 µl 150 kDa Rabbit IgG
Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb 14008 20 µl 155 kDa Rabbit IgG
PLCγ1 (D9H10) XP® Rabbit mAb 5690 20 µl 150 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

PLCγ Antibody Sampler Kit provides an economical means of analyzing phospho and total PLCγ levels. PLCγ Antibody Sampler Kit contains enough primary and secondary antibodies to perform two western blot experiments with each 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

Phosphoinositide-specific phospholipase C (PLC) plays a significant role in transmembrane signaling. In response to extracellular stimuli, such as hormones, growth factors, and neurotransmitters, PLC hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) to generate two secondary messengers: inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DAG) (1). At least four families of PLCs have been identified: PLCβ, PLCγ, PLCδ, and PLCε. Phosphorylation is one of the key mechanisms that regulate the activity of PLC. PLCγ is activated by both receptor and non-receptor tyrosine kinases (2). PLCγ forms a complex with EGF and PDGF receptors, which leads to the phosphorylation of PLCγ at Tyr771, 783, and 1248 (3). Phosphorylation by Syk at Tyr783 activates the enzymatic activity of PLCγ1 (4). PLCγ2 is engaged in antigen-dependent signaling in B cells and collagen-dependent signaling in platelets. Phosphorylation by Btk or Lck at Tyr753, 759, 1197, and 1217 is correlated with PLCγ2 activity (5,6).

  1. Singer, W.D. et al. (1997) Annu Rev Biochem 66, 475-509.
  2. Margolis, B. et al. (1989) Cell 57, 1101-7.
  3. Kim, H.K. et al. (1991) Cell 65, 435-41.
  4. Wang, Z. et al. (1998) Mol Cell Biol 18, 590-7.
  5. Watanabe, D. et al. (2001) J Biol Chem 276, 38595-601.
  6. Ozdener, F. et al. (2002) Mol Pharmacol 62, 672-9.

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.

    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 4
    #3860

    PLCγ Antibody Sampler Kit

    PLCγ Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from serum-starved A431 cells treated with hEGF (100 ng/mL, 5 min.) using Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb #14008. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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.
    PLCγ Antibody Sampler Kit: Image 2 Expand Image
    Simple Western™ analysis of lysates (0.75 mg/ml) from Ramos + IgM cells using Phospho-PLCγ2 (Tyr759) (E9E9Y) Rabbit mAb #50535. The virtual lane view (left) shows the 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 12-230kDa.
    PLCγ Antibody Sampler Kit: Image 3 Expand Image
    Simple Western™ analysis of lysates (1 mg/ml) from ZR-75-1 cells using PLCγ2 (E5U4T) Rabbit mAb #55512. The virtual lane view (left) shows the 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 66-440kDa.
    PLCγ Antibody Sampler Kit: Image 4 Expand Image
    Immunoprecipitation of PLCγ1 protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is PLCγ1 (D9H10) XP® Rabbit mAb, and lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Western blot analysis was performed using PLCγ1 (D9H10) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    PLCγ Antibody Sampler Kit: Image 5 Expand Image
    Simple Western™ analysis of lysates (1mg/mL) from 3T3 cells using PLCγ1 (D9H10) XP® Rabbit mAb #5690. The virtual lane view (left) shows the 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 the12-230 kDa separation module.
    PLCγ Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from NIH/3T3 cells, untreated (-) or stimulated with hPDGF-BB #8912 (5 min; +), and from A-431 cells, untreated (-) or stimulated with hEGF #8916 (5 min; +), using Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    PLCγ Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from Ramos cells, untreated or treated with anti-human IgM, using Phospho-PLCgamma2 (Tyr1217) Antibody (upper) or PLCgamma2 Antibody #3872 (lower).
    PLCγ Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from Daudi cells, untreated (-) or treated with anti-human IgM (12 μg/ml, 10 min; +), using Phospho-PLCγ2 (Tyr759) (E9E9Y) Rabbit mAb (upper), PLCγ2 (E5U4T) Rabbit mAb #55512 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
    PLCγ Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell lines using PLCγ2 (E5U4T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    PLCγ Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from various cell lines using PLCγ1 (D9H10) XP® Rabbit mAb.
    PLCγ Antibody Sampler Kit: Image 11 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.
    PLCγ Antibody Sampler Kit: Image 12 Expand Image
    Immunoprecipitation of phospho-PLCγ1 (Tyr783) from A-431 cell extracts stimulated with hEGF #8916 using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb.
    PLCγ Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of PLCγ2 from HCT 116 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is PLCγ2 (E5U4T) Rabbit mAb. Western blot analysis was performed using PLCγ2 (E5U4T) Rabbit mAb.
    PLCγ Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from HeLa cells, either mock-transfected or transfected for 48 hours with SignalSilence PLCγ1 siRNA I #6293 or siRNA II #6254, using PLCγ1 (D9H10) XP® Rabbit mAb.
    PLCγ Antibody Sampler Kit: Image 15 Expand Image
    Confocal immunofluorescent analysis of HCT 116 cells (left, positive) and 22Rv1 cells (right, negative) using PLCγ2 (E5U4T) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    PLCγ Antibody Sampler Kit: Image 16 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using PLCγ1 (D9H10) XP® Rabbit mAb.
    PLCγ Antibody Sampler Kit: Image 17 Expand Image
    Flow cytometric analysis of NIH/3T3 cells, untreated (blue) or treated with mPDGFbb (200ng/mL, 15 min; green) using Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb (solid lines) or concentration-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.
    PLCγ Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse pancreas using PLCγ1 (D9H10) XP® Rabbit mAb.
    PLCγ Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse spleen using PLCγ1 (D9H10) XP® Rabbit mAb.
    PLCγ Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse testis using PLCγ1 (D9H10) XP® Rabbit mAb.
    PLCγ Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcioma using PLCγ1 (D9H10) XP® Rabbit mAb.