Anti-Phospholipase C gamma 1/PLC-gamma-1 (phospho Y783) 抗体 (ab4828)

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Key features and details

  • Rabbit polyclonal to Phospholipase C gamma 1/PLC-gamma-1 (phospho Y783)
  • Suitable for: IHC-P, WB
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-Phospholipase C gamma 1/PLC-gamma-1 (phospho Y783) antibody
    Phospholipase C gamma 1/PLC-gamma-1 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Phospholipase C gamma 1/PLC-gamma-1 (phospho Y783)

  • 由来種

    Rabbit

  • 特異性

    Phosphorylation site-specific antibody selective for the phosphorylated form of the Phospholipase C gamma 1/PLC-gamma-1 containing a phosphate on tyrosine 783. The antibody has been shown to recognize the endogenous form of Phospholipase C gamma 1/PLC-gamma-1 when phosphorylated on tyrosine 783 in a variety of cell types following treatment by a broad range of extracellular stimuli. Applications include NIH 3T3 cells +/- PDGF. The antibody does not react with a site directed mutant (Y783F).

  • アプリケーション

    適用あり: IHC-P, WBmore details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Rat, Cow, a wide range of other species

  • 免疫原

    Synthetic peptide corresponding to Human Phospholipase C gamma 1/PLC-gamma-1. Derived from the region of Phospholipase C gamma that contains tyrosine 783, based on the human sequence. This region is conserved among many species including human, rat, bovine and frog (X. laevis)
    (Peptide available as ab5249, ab5354)

  • 特記事項

    Previously labelled as Phospholipase C gamma 1. 

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab4828 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-P Use at an assay dependent concentration.
WB 1/1000. Predicted molecular weight: 135 kDa.

ターゲット情報

  • 機能

    Plays a role in actin reorganization and cell migration. The production of the second messenger molecules diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3) is mediated by activated phosphatidylinositol-specific phospholipase C enzymes. Major substrate for heparin-binding growth factor 1 (acidic fibroblast growth factor)-activated tyrosine kinase.

  • 配列類似性

    Contains 1 C2 domain.
    Contains 1 EF-hand domain.
    Contains 2 PH domains.
    Contains 1 PI-PLC X-box domain.
    Contains 1 PI-PLC Y-box domain.
    Contains 2 SH2 domains.
    Contains 1 SH3 domain.

  • ドメイン

    The SH3 domain mediates interaction with CLNK (By similarity). The SH3 domain also mediates interaction with RALGPS1.

  • 翻訳後修飾

    The receptor-mediated activation of PLC-gamma-1 and PLC-gamma-2 involves their phosphorylation by tyrosine kinases in response to ligation of a variety of growth factor receptors and immune system receptors. May be dephosphorylated by PTPRJ.
    Ubiquitinated by CBLB in activated T-cells.

  • 細胞内局在

    Cell projection > lamellipodium. Cell projection > ruffle. Rapidly redistributed to ruffles and lamellipodia structures in response to epidermal growth factor (EGF) treatment.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • 1 phosphatidyl D myo inositol 4 5 bisphosphate antibody
    • 1 phosphatidylinositol 4 5 bisphosphate phosphodiesterase gamma 1 antibody
    • 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1 antibody
    • Inositoltrisphosphohydrolase antibody
    • Monophosphatidylinositol phosphodiesterase antibody
    • NCKAP3 antibody
    • Phosphatidylinositol phospholipase C antibody
    • Phosphoinositidase C antibody
    • Phosphoinositide phospholipase C antibody
    • Phosphoinositide phospholipase C-gamma-1 antibody
    • Phospholipase C 148 antibody
    • Phospholipase C gamma 1 antibody
    • Phospholipase C-gamma-1 antibody
    • Phospholipase C-II antibody
    • PLC gamma 1 antibody
    • PLC II antibody
    • PLC-148 antibody
    • PLC-gamma-1 antibody
    • PLC-II antibody
    • PLC1 antibody
    • PLC148 antibody
    • Plcg1 antibody
    • PLCG1_HUMAN antibody
    • PLCgamma1 antibody
    see all

画像

  • Western blot - Anti-Phospholipase C gamma 1/PLC-gamma-1 (phospho Y783) antibody (ab4828)
    Western blot - Anti-Phospholipase C gamma 1/PLC-gamma-1 (phospho Y783) antibody (ab4828)

    Extracts prepared from NIH/3T3 (Mouse embryonic fibroblast cell line) cells exposed to PDGF (10 ng/mL) for 10 minutes were resolved on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with 1% BSA, followed by incubation with 0.5 µg/mL ab4828 antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected by chemiluminescence using the Tropix WesternStar detection method and Kodak BioMax ultraclear film. The data show that PDGF is a strong inducer of Phospholipase C gamma 1/PLC-gamma-1 phosphorylation on Y783 in NIH3T3 cells. Extracts prepared from NIH-3T3 cells exposed to PDGF (10 ng/mL) for 10 minutes were resolved on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with 1% BSA, followed by incubation with 0.5 µg/mL ab4828 antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected by chemiluminescence using the Tropix

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phospholipase C gamma 1/PLC-gamma-1 (phospho Y783) antibody (ab4828)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phospholipase C gamma 1/PLC-gamma-1 (phospho Y783) antibody (ab4828)

    ab5163 at a diluton of 1/1000 staining Phospholipase C gamma 1/PLC-gamma-1 (phospho Y783) in formalin fixed paraffin embedded sections of a) inflammed human gastric epithelium and b) normal human gastric epithelium by immunohistochemistry. 

参考文献 (1)

ab4828 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab4828 は 1 報の論文で使用されています。

  • Bates RC  et al. Activation of Src and release of intracellular calcium by phosphatidic acid during Xenopus laevis fertilization. Dev Biol N/A:N/A (2013). Xenopus laevis . PubMed: 24269904

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Question

DESCRIPTION OF THE PROBLEM weak signals, therefore very low signal-background ratio SAMPLE rat, endothelial cells from heart, whole cell lysate PRIMARY ANTIBODY TBS containing 0.1% Tween, 5% BSA overnight at 4°C with ab4828 1:1000, 1:500, 1:200 DETECTION METHOD ECL Plus POSITIVE AND NEGATIVE CONTROLS USED As positive controls: 2h 100ng PMA and 100µM ATP for 2 min not a real negative control: unstimulated cells ANTIBODY STORAGE CONDITIONS aliquot, -24°C SAMPLE PREPARATION Cell lysis: PBS w/o calcium, w/o magnesium; 1% NP-40; 1mM PMSF, 1µg/ml Pepstatin, 40mM beta-Glycerophosphate, Sigma Phosphatase Cocktail 2 mechanical destruction and sonification at 4°C Sample Preparation according to invitrogen/Nupage gels LDS Sample buffer, reducing agent, 10min 70°C heating AMOUNT OF PROTEIN LOADED approx 150µg/band ELECTROPHORESIS/GEL CONDITIONS NuPage Electrophoresis System, 4-12% Bis-Tris Mini-Gel, MOPS buffer, reducing agent, 200V constant 50 min TRANSFER AND BLOCKING CONDITIONS PVDF membrane, Tansferbuffer Invitrogen, 1h 40 min 20V Ponceau TBS containing 0.1% Tween, 5% non-fat dry milk for 1 h 2x2min washing in TBS containing 0.1% Tween SECONDARY ANTIBODY TBS containing 0.1% Tween, 5% non-fat dry milk for 1 h (other company)ECL anti-rab IgG (NA934V) 1:4000 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 6 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? primary antibody concentration (up to 1:200) and protein loading (maximum for these commercial gels) ADDITIONAL NOTES I could detect PLC phosphorylation at tyr 771 with the identical protocol (good signal to background level). But just get really weak signals just below background with the tyr 783, and I only get these signals when I increase the detection time up to 20 min (!). (Biorad CCD detection system) I would like to know if there is a treatment/stimulation for my endothelial cells which causes a phosphorylation at tyr 783 without any doubt.

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Answer

Thank you for your enquiry. I am sorry to hear you are having a problem with ab4828 (Phospholipase C gamma 1 (phospho Y783) antibody). The following information from the datasheet should help improve your results: Extracts prepared from NIH-3T3 cells exposed to PDGF (10 ng/mL) for 10 minutes were resolved on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with 1% BSA, followed by incubation with 0.5 µg/mL ab4828 antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected by chemiluminescence using the Tropix WesternStar detection method and Kodak BioMax ultraclear film. The data show that PDGF is a strong inducer of PLC gamma 1 phosphorylation on Y783 in NIH3T3 cells. The WB image is available on the ab4828 datasheet. Please let me know if the above suggestions improve your results. Note that it is important to include the NIH-3T3 cells as a positive control. I look forward to your reply.

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Question

Customer would like to know the concentration.

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Answer

The originator of ab4828 was able to give me a concentration for this antibody - 0.25 mg/mL. Please let me know if you have any more questions.

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Question

Customer would like to know the concentration.

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Answer

Thank you for your enquiry. There isn't a concentration for this glycerol format antibody. Ab4828 was packaged based on volume in order to be able to adjust the concentration of the antibody to assure consistent batch-to-batch signal strength using the originator's prequalified test systems. Therefore the actual amount of antibody is less critical than the signal intensity provided. For Western blotting it is recommended to use ab4828 at a 1:1000 dilution. If you have any more questions please contact us again.

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