Cell Signaling Technology

Product Pathways - Ca / cAMP / Lipid Signaling

Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb #8713

No. Size Price
8713S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
8713 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Monkey, Endogenous 150 Rabbit IgG
IP 1:50
IHC-P 1:200
F 1:200
IF-IC 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Rat,

Specificity / Sensitivity

Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb recognizes endogenous levels of PLCγ1 protein only when phosphorylated at Ser1248.

Phospho-PLCγ1(Ser1248) (D25A9)兔单克隆抗体可识别内源性的Ser1248磷酸化的PLCγ1。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser1248 of human PLCγ1 protein.

该单克隆抗体用与人类PLCγ1蛋白中Ser1248位点附近的氨基酸序列对应的人工合成肽段免疫动物而制成。

IF-IC

IF-IC

Confocal immunofluorescent analysis of A-431 cells, serum starved (left) or treated with hEGF #8916 (100 ng/mL for 15 min) using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

对A-431细胞,血清饥饿处理(左)或100ng/mL hEGF #8916处理15分钟(右),使用Phospho-PLCγ1(Ser1248)(D25A9)兔单克隆抗体(绿色)进行共聚焦免疫荧光分析。蓝色伪彩=DRAQ5®#4084(荧光DNA染料)。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, treated with U0126 #9903 (blue) or TPA #4174 (green), using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb.

对Jurkat细胞,经U0126 #9903(蓝色)或TPA #4174(绿色)处理,使用Phospho-PLCγ1(Ser1248) (D25A9)兔抗体进行流式细胞仪分析。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of SignalSlide® Phospho-EGF Receptor IHC Controls #8102 [paraffin-embedded KYSE450 cell pellets untreated (left) or EGF-treated (right)] using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb.

对石蜡包埋的KYSE450细胞,未处理(左)或EGF处理(右),使用Phospho-PLCγ1(Ser1248) (D25A9)兔单抗进行SignalSlide Phospho-EGF Receptor IHC Controls #8102免疫组化分析。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb.

对石蜡包埋的人乳腺癌使用Phospho-PLCγ1(Ser1248) (D25A9)兔单抗进行免疫组化分析。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon (normal adjacent to tumor) using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

对石蜡包埋的人结肠(癌旁正常组织)使用Phospho-PLCγ1(Ser1248) (D25A9)兔单抗进行免疫组织化学分析,其中左图使用对照肽处理,右图使用抗原特异肽处理。

Western Blotting

Western Blotting

Western blot analysis of extracts from serum-starved A-431 and A549 cells, untreated (-) or treated (+) with hEGF #8916 (100 ng/mL, 15 min) or serum-starved NIH/3T3 cells, untreated (-) or treated (+) with hPDGF-BB #8912 (50 ng/mL, 15 min), using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb (upper) or PLCγ1 (D9H10) XP® Rabbit mAb #5690 (lower).

对血清饥饿的A431和A549细胞抽提液,未处理(-)或100ng/ml hEGF #8916处理15分钟(+),或者是血清饥饿的NIH/3T3细胞抽提液,未处理(-)或50ng/ml hPDGF-BB #8912处理15分钟(+),使用Phospho-PLCγ1(Ser1248) (D25A9)(上图)或PLCγ1 (D9H10) XP? 兔单抗 #5690(下图)进行Western blot分析。

IP

IP

Immunoprecipitation (IP)/Western blot analysis of extracts from serum-starved HeLa cells, untreated (-) or treated (+) with TPA #4174 (100 nM, 15 min) prior to lysis in SDS (lanes 1 and 2) or IP lysis buffer (lane 3, TPA-treated only). IP Lysates were then subjected to immunoprecipitation with Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb (lane 4), PLCγ1 (D9H10) XP® Rabbit mAb #5690 (lane 5), or Normal Rabbit IgG #2729 (lane 6). The western blot was probed using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb. Lane 3 represents 10% input.

对血清饥饿的HeLa细胞,未处理(-)或用100nM TPA #4174 (+) 处理15分钟后,用SDS裂解(列1,2)或IP裂解缓冲液裂解(列3,仅TPA处理),然后进行免疫沉淀(IP)/Western blot分析。IP裂解物用Phospho-PLCγ1(Ser1248) (D25A9)兔单抗(列4)、PLCγ1 (D9H10) XP®兔单抗 #5690(列5),或正常鼠IgG #2729(列6)进行免疫沉淀。Western blot分析使用Phospho-PLCγ1 (Ser1248) (D25A9)兔单抗。列3代表了10%的输入对照。

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 1245 (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).

磷酸肌醇特异性磷脂酶C(PLC)在跨膜信号转导中扮演明显的作用。在对激素、生长因子和神经递质反应的过程中,PLC水解4,5-二磷酸磷酸酰肌醇,产生两个第二信使:1,4,5-三磷酸肌醇(IP3)和甘油二酯(DAG)(1)。至少四个PLC家族已被鉴定,分别为PLC-α、PLC-β、PLC-γ和PLC-δ。磷酸化是调节PLC活性的重要机制之一。PLCγ被受体和非受体酪氨酸激酶激活(2)。PLCγ与EGF和PDGF受体形成复合物,导致PLCγ的Tyr771、783和1245位点磷酸化(3)。Syk对Tyr783的磷酸化激活PLCγ1的酶活性(4)。PLCγ2参与B细胞抗原依赖的信号转导和血小板胶原依赖的信号转导。Btk或Lck对Tyr753、759、1197和1217的磷酸化与PLCγ2的活性相关(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.
  7. Burgess, W.H. et al. (1990) Mol Cell Biol 10, 4770-7.
  8. Ohta, S. et al. (1988) FEBS Lett 242, 31-5.
  9. Rodriguez, R. et al. (2001) J Biol Chem 276, 47982-92.
  10. Humphries, L.A. et al. (2004) J Biol Chem 279, 37651-61.
  11. Kim, Y.J. et al. (2004) Mol Cell Biol 24, 9986-99.
  12. Sekiya, F. et al. (2004) J Biol Chem 279, 32181-90.
  13. Carpenter, G. and Ji, Q. (1999) Exp Cell Res 253, 15-24.
  14. Rebecchi, M.J. and Pentyala, S.N. (2000) Physiol Rev 80, 1291-335.
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  16. Wang, Y. et al. (2006) Mol Biol Cell 17, 2267-77.
  17. Park, D.J. et al. (1992) J Biol Chem 267, 1496-501.

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