Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20920-19] to MPTX+MPTX2
  • Suitable for: WB, IHC-P, IHC-Fr, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse

Overview

  • Product name

    Anti-MPTX+MPTX2 antibody [EPR20920-19]
    See all MPTX+MPTX2 primary antibodies

  • Description

    Rabbit monoclonal [EPR20920-19] to MPTX+MPTX2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, ICC/IF, Flow Cytmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: His-tagged human MPTX recombinant protein. His-lif tagged human MPTX2 recombinant protein. Wild type mouse large intestine tissue lysate. IHC-Fr: Mouse small intestine tissue. IHC-P: Mouse small intestine tissue. ICC/IF: HEK-293T cells transfected with a flag-tagged MPTX expression construct.Flow Cyt: HEK-293T cells transfected with a flag-tagged MPTX expression construct.

  • 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR20920-19

  • Isotype

    IgG

Images

  • Immunohistochemistry (Frozen sections) - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)
    Immunohistochemistry (Frozen sections) - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)

    Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen mouse small intestine tissue labeling MPTX1 with ab238123 at 1/500 (1.10 µg/ml) dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 µg/ml) dilution. The nuclear counterstain was DAPI (Blue). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 µg/ml) dilution.

    Positive staining on Paneth cells in mouse small intestine is observed (PMID: 29144463).

  • Western blot - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)
    Western blot - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)
    All lanes : Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123) at 1/1000 dilution

    Lane 1 : Wild type mouse large intestine tissue lysate
    Lane 2 : MPTX knockout mouse large intestine tissue lysate

    Lysates/proteins at 10 µg per lane.

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

    Observed band size: 27 kDa
    why is the actual band size different from the predicted?



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

    MPTX2 is reported to express in Paneth cells in the small intestine (PMID: 29144463).

    The lysates were kindly provided by an anonymous collaborator.

    ab238123 was shown to specifically react with MPTX in wild-type mouse intestine tissue  as signal was lost in MPTX knockout tissue. Wild-type and MPTX knockout samples were subjected to SDS-PAGE. ab238123 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ instrument using the ECL technique.  Exposure time: 15 seconds.

  • Western blot - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)
    Western blot - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)
    All lanes : Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123) at 1/1000 dilution

    Lane 1 : His-tagged human MPTX recombinant protein, 10ng
    Lane 2 : His-lif tagged human MPTX2 recombinant protein, 10ng

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

    Observed band size: 27/75 kDa why is the actual band size different from the predicted?



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

    MPTX2 protein carries a lif tag (MW around 40 kDa). 

    Exposure time: 3 seconds.

  • Flow Cytometry - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)
    Flow Cytometry - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (human embryonic kidney epithelial cell) transfected with a flag-tagged MPTX expression construct cells labeling MPTX1 with ab238123 at 1/500 (Right panel) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left panel). A Goat anti rabbit IgG (Alexa Fluor® 647, ab150079) at 1/2000 was used as the secondary antibody.

  • Immunocytochemistry/ Immunofluorescence - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)
    Immunocytochemistry/ Immunofluorescence - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) transfected with a flag-tagged MPTX expression construct cells labeling MPTX1 with ab238123 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing positive staining in HEK-293T cells transfected with a flag-tagged MPTX expression construct.

    Mouse monoclonal anti-DDDDK antibody followed by ab150120 AlexaFluor®594 Goat anti-Mouse secondary was used to counterstain at 1/500 dilution (Red). The nuclear counterstain was DAPI (Blue).

    -ve control 1: ab238123 at a 1/50 dilution followed by ab150120 at a 1/1000 dilution.

    -ve control 2: Mouse monoclonal anti-DDDDK antibody at a 1/500 dilution followed by ab150077 at a 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)

    Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue labeling MPTX1 with ab238123 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on Paneth cells of mouse small intestine is observed. The section was incubated with ab238123 for 5 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

    Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins.

  • Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)
    Anti-MPTX+MPTX2 antibody [EPR20920-19] (ab238123)

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