Recombinant
RabMAb

Recombinant Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)

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Overview

  • Product name

    Anti-PHAPI2 / APRIL antibody [EPR14588]
    See all PHAPI2 / APRIL primary antibodies

  • Description

    Rabbit monoclonal [EPR14588] to PHAPI2 / APRIL

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, Flow Cyt, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human PHAPI2/ APRIL aa 1-100. The exact sequence is proprietary.
    Database link: Q92688

  • Positive control

    • WB: PC-3, LNCap, Jurkat, C6, RAW 264.7, PC-12, and NIH/3T3 whole cell lysates; Human fetal heart and fetal kidney lysates. IHC-P: Human prostatic hyperplasia, Human spleen, mouse spleen and rat cardiac muscle tissues. ICC/IF: PC-3 and Jurkat cells. Flow Cyt: Jurkat cells.

  • 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

Applications

Our Abpromise guarantee covers the use of ab200836 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
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
Flow Cyt 1/180.
WB 1/1000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).

Target

  • Function

    Multifunctional protein working as a cell cycle progression factor as well as a cell survival factor. Required for the progression from the G1 to the S phase. Anti-apoptotic protein which functions as a caspase-3 inhibitor. Has no phosphatase 2A (PP2A) inhibitor activity (By similarity). Exhibits histone chaperone properties, stimulating core histones to assemble into a nucleosome.

  • Tissue specificity

    Expressed in heart, lung, pancreas, prostate and in spleen, thymus and placenta.

  • Sequence similarities

    Belongs to the ANP32 family.
    Contains 4 LRR (leucine-rich) repeats.
    Contains 1 LRRCT domain.

  • Domain

    Histone binding is mediated by the concave surface of the LRR region.

  • Post-translational
    modifications

    Some glutamate residues are glycylated by TTLL8. This modification occurs exclusively on glutamate residues and results in a glycine chain on the gamma-carboxyl group.

  • Cellular localization

    Nucleus. Accumulates in the nuclei at the S phase and Cytoplasm. Lacks a nuclear localization signal.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Acidic (leucine rich) nuclear phosphoprotein 32 family member B antibody
    • Acidic leucine rich nuclear phosphoprotein 32 family member B antibody
    • Acidic leucine-rich nuclear phosphoprotein 32 family member B antibody
    • Acidic nuclear phosphoprotein 32 family member B antibody
    • Acidic protein rich in leucines antibody
    • AN32B_HUMAN antibody
    • ANP 32B antibody
    • ANP32B antibody
    • APRIL antibody
    • OTTHUMP0000006377 antibody
    • PAL 31 antibody
    • PAL31 antibody
    • PHAPI 2 antibody
    • PHAPI2 antibody
    • PHAPI2 protein antibody
    • PHAPI2a antibody
    • Proliferation related acidic leucine rich protein antibody
    • Putative HLA-DR-associated protein I-2 antibody
    • Silver stainable protein SSP 29 antibody
    • Silver stainable protein SSP29 antibody
    • Silver-stainable protein SSP29 antibody
    • Ssp 29 antibody
    • Ssp29 antibody
    see all

Images

  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836) at 1/1000 dilution + PC-3 (Human prostate cancer cell line) whole cell lysate at 10 µg

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

    Predicted band size: 29 kDa
    Observed band size: 29 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Immunocytochemistry/ Immunofluorescence - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-3 (Human prostate cancer cell line) cells labeling PHAPI2 / APRIL with ab200836 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear and cytoplasmic staining on PC-3 cell line is observed. The nuclear counter stain 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 1/500 dilution (red).

  • Immunocytochemistry/ Immunofluorescence - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Immunocytochemistry/ Immunofluorescence - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling PHAPI2 / APRIL with ab200836 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear and cytoplasmic staining on Jurkat cell line is observed. The nuclear counter stain 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 1/500 dilution (red).
    The negative controls are as follows;
    -ve control 1: ab200836 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 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 1/500 dilution.

     

  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    All lanes : Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836) at 1/10000 dilution

    Lane 1 : LNCaP (Human prostate cancer cell line) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) 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: 29 kDa
    Observed band size: 29 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    All lanes : Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836) at 1/1000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 29 kDa
    Observed band size: 29 kDa


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    All lanes : Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : RAW 264.7 (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: 29 kDa
    Observed band size: 29 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    All lanes : Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836) at 1/1000 dilution

    Lane 1 : PC-3 (Human prostate cancer cell line) whole cell lysate
    Lane 2 : PC-3 (Human prostate cancer cell line) whole cell lysate with PHAPI2 / APRIL peptide
    Lane 3 : PC-3 (Human prostate cancer cell line) whole cell lysate with PHAPI peptide

    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: 29 kDa
    Observed band size: 29 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Based on sequence analysis ab200836 has 78% homology with PHAPI protein. The levels of XR were tested in the accompanying blocking experiment.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)

    Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling PHAPI2 / APRIL with ab200836 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human prostatic hyperplasia tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)

    Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling PHAPI2 / APRIL with ab200836 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human spleen tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling PHAPI2 / APRIL with ab200836 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Mouse spleen tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)

    Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling PHAPI2 / APRIL with ab200836 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Rat cardiac muscle tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Flow Cytometry - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
    Flow Cytometry - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)

    Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling PHAPI2 / APRIL with ab200836 at 1/180 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

References

ab200836 has not yet been referenced specifically in any publications.

Publishing research using ab200836? Please let us know so that we can cite the reference in this datasheet.

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