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    phapi2--april-antibody-epr14588-ab200836.pdf

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Cell Biology Apoptosis Extracellular Signals Death Ligands
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Validated using a knockout cell lineRecombinantRabMAb

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

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Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Immunocytochemistry/ Immunofluorescence - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Immunocytochemistry/ Immunofluorescence - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • 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)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Flow Cytometry (Intracellular) - Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)
  • Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR14588] to PHAPI2 / APRIL
  • Suitable for: IHC-P, WB, Flow Cyt (Intra), ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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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, WB, Flow Cyt (Intra), ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK293T, 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: Raw264.7 and Jurkat cells. Flow Cyt (intra): Jurkat cells.
  • 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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR14588
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Extracellular Signals
    • Death Ligands
    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Inhibitors
    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Other cell division antibodies

Associated products

  • Alternative Versions

    • Anti-PHAPI2 / APRIL antibody [EPR14588] - BSA and Azide free (ab239014)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human ANP32B (PHAPI2/APRIL) knockout HEK-293T cell line (ab266818)
  • KO cell lysates

    • Human ANP32B (PHAPI2/APRIL) knockout HEK-293T cell lysate (ab257831)
  • Recombinant Protein

    • Recombinant Human PHAPI2 / APRIL protein (ab133003)

Applications

The Abpromise guarantee

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

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.
  • Target information above from: UniProt accession Q92688 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10541 Human
    • Entrez Gene: 67628 Mouse
    • Entrez Gene: 170724 Rat
    • SwissProt: Q92688 Human
    • SwissProt: Q9EST5 Mouse
    • SwissProt: Q9EST6 Rat
    • Unigene: 494604 Human
    • Unigene: 263913 Mouse
    • Unigene: 20465 Rat
    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)
    All lanes : Anti-PHAPI2 / APRIL antibody [EPR14588] (ab200836) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : ANP32B knockout HEK293T cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Lncap cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

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



    Lanes 1-4: Merged signal (red and green). Green - ab200836 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab200836 Anti-PHAPI2 / APRIL antibody [EPR14588] was shown to specifically react with PHAPI2 / APRIL in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266818 (knockout cell lysate ab257831) was used. Wild-type and PHAPI2 / APRIL knockout samples were subjected to SDS-PAGE. ab200836 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • 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.

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

  • 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 Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling PHAPI2 / APRIL with ab200836 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Ab195889 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/200 dilution (red). Confocal image showing mostly nuclear staining in Raw264.7 cell line.

  • 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.

  • 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.

  • 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.

     

  • 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.

    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-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.

    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-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.

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

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

    Intracellular 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. 

     

     

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

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (0)

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ab200836 has not yet been referenced specifically in any publications.

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