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

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: ICC, IHC-P, WB, Flow Cyt (Intra)more 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.

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

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)

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
ICC		1/500.
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.

Notes
ICC 1/500.
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.

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

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)

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 - 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)

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)

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)

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)

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)

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 - 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)

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)

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)

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)

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)

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

Click here to view the general protocols

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Key features and details

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

Post-translational
modifications