Recombinant
RabMAb

Recombinant Anti-Phospholipase C beta 1 + Phospholipase C beta 3 antibody [EPR18714] (ab184743)

Overview

  • Product name

    Anti-Phospholipase C beta 1 + Phospholipase C beta 3 antibody [EPR18714]
    See all Phospholipase C beta 1+Phospholipase C beta 3 primary antibodies
  • Description

    Rabbit monoclonal [EPR18714] to Phospholipase C beta 1 + Phospholipase C beta 3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Human Phospholipase C beta 3/PLCB3 aa 550-750. Cross reacts with Phospholipase C beta 1 (Q9Z1B3).
    Database link: Q01970

  • Positive control

    • WB: Human Phospholipase C beta 1 and beta 3 fragment recombinant proteins; Mouse and Rat cerebellum and brain lysates; Human fetal brain lysate; HCT 116, HEK-293, U-87 MG and SH-SY5Y whole cell lysates. IHC-P: Human cerebrum, Mouse cerebellum and Rat cerebellum tissues. IP: Mouse brain whole cell lysate.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18714
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab184743 in the following tested applications.

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

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 150 kDa (predicted molecular weight: 139 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/20.

Target

Images

  • All lanes : Anti-Phospholipase C beta 1 + Phospholipase C beta 3 antibody [EPR18714] (ab184743) at 1/5000 dilution

    Lane 1 : Human Phospholipase C beta 1 fragment recombinant protein
    Lane 2 : Human Phospholipase C beta 3 fragment recombinant protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 139 kDa


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human Phospholipase C beta 1 recombinant protein fragment contains aa518-656 with a His-Tag®. Human Phospholipase C beta 3 recombinant protein fragment contains aa590-706 with a His-Tag®. The immunogen of this product has low homology with Phospholipase C beta 2 and Phospholipase C beta 4.

  • All lanes : Anti-Phospholipase C beta 1 + Phospholipase C beta 3 antibody [EPR18714] (ab184743) at 1/1000 dilution

    Lane 1 : Mouse cerebellum lysate
    Lane 2 : Rat cerebellum lysate
    Lane 3 : Human fetal brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lane 3 : Goat Anti-Rabbit IgG Peroxidase conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

    Predicted band size: 139 kDa
    Observed band size: 150 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.
    The molecular weight observed is consistent with what has been described in the literature (PMID: 8429045, 17298601). Phospholipase C beta 1 selectively expresses in the brain which has been described in the literature (PMID: 2468162).

  • All lanes : Anti-Phospholipase C beta 1 + Phospholipase C beta 3 antibody [EPR18714] (ab184743) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat heart lysate
    Lane 7 : Rat kidney lysate
    Lane 8 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

    Predicted band size: 139 kDa
    Observed band size: 150 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1,2,3 & 4: 30 seconds; Lane 5,6,7 & 8:10 seconds.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 8429045, 17298601). Phospholipase C beta 1 selectively expresses in the brain which has been described in the literature (PMID: 2468162).

  • All lanes : Anti-Phospholipase C beta 1 + Phospholipase C beta 3 antibody [EPR18714] (ab184743) at 1/1000 dilution

    Lane 1 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
    Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 139 kDa
    Observed band size: 150 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Phospholipase C beta 1 + Phospholipase C beta 3 with ab184743 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Mainly cytoplasm staining on neurons of the normal Human cerebrum is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling Phospholipase C beta 1 + Phospholipase C beta 3 with ab184743 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on purkinje cells of the Mouse cerebellum is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling Phospholipase C beta 1 + Phospholipase C beta 3 with ab184743 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on purkinje cell of the Rat cerebellum is observed. 

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Phospholipase C beta 1 + Phospholipase C beta 3 with ab184743 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Human kidney. Counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Phospholipase C beta 1 + Phospholipase C beta 3 with ab184743 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Mouse liver. Counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Phospholipase C beta 1 + Phospholipase C beta 3 with ab184743 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Rat testis. Counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Phospholipase C beta 1 + Phospholipase C beta 3 were immunoprecipitated from 1mg of Mouse brain whole cell lysate with ab184743 at 1/20 dilution.

    Western blot was performed from the immunoprecipitate using ab184743 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Mouse brain whole cell lysate, 10ug (Input).

    Lane 2: ab184743 IP in Mouse brain whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab184743 in Mouse brain whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

References

ab184743 has not yet been referenced specifically in any publications.

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