Recombinant
RabMAb

Recombinant Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

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Overview

  • Product name

    Anti-PPCEL/PREPL antibody [EPR17752]
    See all PPCEL/PREPL primary antibodies

  • Description

    Rabbit monoclonal [EPR17752] to PPCEL/PREPL

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human PPCEL/PREPL aa 600-700. The exact sequence is proprietary.
    Database link: Q4J6C6

  • Positive control

    • WB: Human fetal brain, MCF-7, Mouse brain, Rat brain, C6, Raw264.7, PC-12 or NIH/3T3 whole cell lysate. IHC-P: Human, Mouse or Rat cerebral cortex tissue. ICC/IF: SH-SY5Y or MCF-7 cells. FC: MCF-7 cells.

  • General notes

     

     

     This product was previously labelled as PPCEL

     

    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

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR17752

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab203111 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 1/2000. Detects a band of approximately 72 kDa (predicted molecular weight: 84 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/250.
IP 1/30.

Target

  • Function

    Probable serine peptidase whose precise substrate specificity remains unclear. Does not cleave peptides after a arginine or lysine residue.

  • Tissue specificity

    Widely expressed. Expressed at higher level in brain, skeletal muscle, heart and kidney.

  • Involvement in disease

    Defects in PREPL are a cause of hypotonia-cystinuria syndrome (HCS) [MIM:606407]. HCS is characterized generalized hypotonia at birth, nephrolithiasis, growth hormone deficiency, minor facial dysmorphism, failure to thrive, followed by hyperphagia and rapid weight gain in late childhood. HCS is caused by a deletion that disrups both SLC3A1 and PREPL genes. As SLC3A1 is known to cause isolated cystinuria type I, the extended phenotype could be attributed to the deletion of PREPL.

  • Sequence similarities

    Belongs to the peptidase S9A family.

  • Cellular localization

    Cytoplasm > cytosol.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • PPCEL_HUMAN antibody
    • prepl antibody
    • Prolyl endopeptidase-like antibody
    • Prolylendopeptidase-like antibody
    • putative prolyl oligopeptidase antibody
    see all

Images

  • Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Lane 1 : Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/1000 dilution
    Lane 2 : Anti-GST antibody [EPR4236] (ab111947) at 1/10000 dilution

    All lanes : Recombinant Human PPCEL/PREPL protein (ab160557)

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 84 kDa
    Observed band size: 109 kDa
    why is the actual band size different from the predicted?



    Exposure time:

    Lane 1: 3 seconds.

    Lane 2: 3 minutes.

     

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Immunocytochemistry/ Immunofluorescence - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling PPCEL/PREPL with ab203111 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on SH-SY5Y cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:-
    -ve control 1 - ab203111 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Immunocytochemistry/ Immunofluorescence - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF-7(Human breast adenocarcinoma cell line) cells labeling PPCEL/PREPL with ab203111 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on MCF-7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:-
    -ve control 1 - ab203111 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    All lanes : Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/2000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : MCF-7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 84 kDa
    Observed band size: 72 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution buffer 5% NFDM/TBST.

     

    The observed MW is consistent with what has been described in the literature PMID: 16385448.

  • Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    All lanes : Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/2000 dilution

    Lane 1 : Mouse brain whole cell lysate
    Lane 2 : Rat brain whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 84 kDa
    Observed band size: 72 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/DIlution buffer 5% NFDM/TBST.

     

    The observed MW is consistent with what has been described in the literature PMID: 16385448.

  • Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    All lanes : Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 84 kDa
    Observed band size: 72 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer 5% NFDM/TBST.

     

    The observed MW is consistent with what has been described in the literature PMID: 16385448.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

    Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling PPCEL/PREPL with ab203111 at 1/250 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a 1/500 dilution. Cytoplasmic staining on Human cerebral cortex tissue is observed. Counter stained with Hematoxylin.

     

    Negative control: Used PBS instead of primary antibody, secondary antibody is ab97051 Goat Anti-Rabbit IgG H&L (HRP).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

    Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling PPCEL/PREPL with ab203111 at 1/250 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a 1/500 dilution. Cytoplasmic staining on Mouse cerebral cortex tissue is observed. Counter stained with Hematoxylin.

     

    Negative control: Used PBS instead of primary antibody, secondary antibody is ab97051 Goat Anti-Rabbit IgG H&L (HRP).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

    Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling PPCEL/PREPL with ab203111 at 1/250 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a 1/500 dilution. Cytoplasmic staining on Rat cerebral cortex tissue is observed. Counter stained with Hematoxylin.

     

    Negative control: Used PBS instead of primary antibody, secondary antibody is ab97051 Goat Anti-Rabbit IgG H&L (HRP).

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

  • Immunoprecipitation - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)
    Immunoprecipitation - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

    PPCEL/PREPL was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell extract with ab203111 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab203111 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: NIH/3T3 whole cell extract 10 µg (Input). Lane 2: ab203111 IP in NIH/3T3 whole cell extract. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab203111 in NIH/3T3 whole cell extract.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds

     

    Blocking/Dilution Buffer: 5% NFDM/TBST

References

ab203111 has not yet been referenced specifically in any publications.

Publishing research using ab203111? Please let us know so that we can cite the reference in this datasheet.

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