Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Pit1 antibody [EPR23555-203] (ab273048)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23555-203] to Pit1
  • Suitable for: WB, Flow Cyt, IP, IHC-P, IHC-Fr, ICC/IF
  • Reacts with: Mouse, Rat

Overview

  • Product name

    Anti-Pit1 antibody [EPR23555-203]
    See all Pit1 primary antibodies

  • Description

    Rabbit monoclonal [EPR23555-203] to Pit1

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Rat
    ICC/IF
    Rat
    IHC-Fr
    Mouse
    IHC-P
    Mouse
    IP
    Mouse
    Rat
    WB
    Mouse
    Rat
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: GH3 whole cell lysate; Mouse pituitary tissue. IHC-P: Mouse pituitary tissue. IHC-Fr: Mouse pituitary tissue. ICC/IF: GH3 cells. Flow Cyt: GH3 cells. IP: Mouse pituitary tissue lysate. GH3 whole cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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

    EPR23555-203

  • Isotype

    IgG

  • Research areas

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)

    Immunohistochemical analysis of paraffin-embedded mouse pituitary tissue labeling Pit1 with ab273048 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in mouse pituitary (PMID:21123951). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Frozen sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Immunohistochemistry (Frozen sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse pituitary tissue labeling Pit1 with ab273048 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2µg/ml) dilution (Green). Nuclear staining on mouse pituitary is observed. The nuclear counterstain was DAPI (Blue).

    Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2µg/ml) dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunoprecipitation - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Immunoprecipitation - Anti-Pit1 antibody [EPR23555-203] (ab273048)

    Pit1 was immunoprecipitated from 0.35 mg mouse pituitary tissue lysate 10µg with ab273048 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab273048 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: Mouse pituitary tissue lysate 10µg.

    Lane 2: ab273048 IP in mouse pituitary tissue lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab273048 in mouse pituitary tissue lysate.

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

    Exposure time: 23 seconds.

     

  • Western blot - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Western blot - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    All lanes : Anti-Pit1 antibody [EPR23555-203] (ab273048) at 1/1000 dilution

    Lane 1 : Mouse pituitary tissue lysate
    Lane 2 : Mouse liver tissue lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 33 kDa
    Observed band size: 33 kDa


    Exposure time: 81 seconds


    The expression profile observed is consistent with what has been described in the literature (PMID: 21123951).
    Negative control: Mouse liver (PMID: 21123951).

    Blocking/Diluting buffer and concentration 5% NFDM/TBST

  • Western blot - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Western blot - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Anti-Pit1 antibody [EPR23555-203] (ab273048) at 1/1000 dilution + GH3 (Rat pituitary epithelial), whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 33 kDa
    Observed band size: 31,33 kDa
    why is the actual band size different from the predicted?



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

    The expression profile observed is consistent with what has been described in the literature. The 31kDa and 33kDa bands correspond to different isoforms (PMID: 21123951, 21060149).

    Exposure time: 7 seconds.

  • Immunocytochemistry/ Immunofluorescence - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Immunocytochemistry/ Immunofluorescence - Anti-Pit1 antibody [EPR23555-203] (ab273048)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized GH3 (Rat pituitary epithelial cell line) cells labelling Pit1 with ab273048 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2µg/ml dilution (Green). Confocal image showing nuclear staining in GH3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5µg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2µg/ml dilution.

  • Flow Cytometry - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Flow Cytometry - Anti-Pit1 antibody [EPR23555-203] (ab273048)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized GH3 (Rat pituitary epithelial cell line) cells labelling Pit1 with ab273048 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Immunoprecipitation - Anti-Pit1 antibody [EPR23555-203] (ab273048)

    Pit1 was immunoprecipitated from 0.35 mg GH3 (Rat pituitary epithelial), whole cell lysate 10µg with ab273048 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab273048 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: GH3 (Rat pituitary epithelial), whole cell lysate 10µg.

    Lane 2: ab273048 IP in GH3 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab273048 in GH3 whole cell lysate.

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

    Exposure time: 5 seconds.

  • Immunohistochemistry (Frozen sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Immunohistochemistry (Frozen sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling Pit1 with ab273048 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2µg/ml) dilution (Green). No staining on mouse liver is observed. The nuclear counterstain was DAPI (Blue).

    Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2µg/ml) dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)

    Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Pit1 with ab273048 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with Hematoxylin.

    Negative control: No staining in mouse cerebrum (PMID:21123951).

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Anti-Pit1 antibody [EPR23555-203] (ab273048)
    Anti-Pit1 antibody [EPR23555-203] (ab273048)

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