Anti-PHAP1 antibody (ab5991)
Key features and details
- Rabbit polyclonal to PHAP1
- Suitable for: WB, ICC/IF, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-PHAP1 antibody
See all PHAP1 primary antibodies -
Description
Rabbit polyclonal to PHAP1 -
Host species
Rabbit -
Specificity
This antibody does not cross react with PHAP12a or PHAPIII isoforms. -
Tested applications
Suitable for: WB, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human PHAP1 aa 199-249 (C terminal). The exact sequence is proprietary. A peptide corresponding to 14 amino acids near the carboxy terminus of human PHAP1.
(Peptide available asab6240) -
Positive control
- WB: Wild -type HEK-293T cell lysate. THP-1 and HeLa cell lysate.
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General notes
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Primary antibody notes
Apoptosis is related to many diseases and development. Caspase-9 plays a central role in cell death induced by a variety of apoptosis activators. Cytochrome c, after released from mitochondria, binds to Apaf-1, which forms an apoptosome that in turn binds to and activate procaspase-9. Activated caspase-9 cleaves and activates the effector caspases (caspase-3, -6 and –7), which are responsible for the proteolytic cleavage of many key proteins in apoptosis. The tumor suppressor putative HLA-DR-associated proteins (PHAPs) were recently identified as important regulators of mitochondrion apoptosis. PHAP appears to facilitate apoptosome-mediated caspase-9 activation and to stimulate the mitochondrial apoptotic pathway. PHAP was also shown to oppose both Ras- and Myc-mediated cell transformation. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab5991 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 |
|---|---|---|
| WB |
Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 32 kDa.
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| ICC/IF |
Use a concentration of 20 µg/ml.
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| IHC-P |
Use a concentration of 2 µg/ml.
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| Notes |
|---|
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WB
Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 32 kDa. |
|
ICC/IF
Use a concentration of 20 µg/ml. |
|
IHC-P
Use a concentration of 2 µg/ml. |
Target
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Function
Implicated in a number of cellular processes, including proliferation, differentiation, caspase-dependent and caspase-independent apoptosis, suppression of transformation (tumor suppressor), inhibition of protein phosphatase 2A, regulation of mRNA trafficking and stability in association with ELAVL1, and inhibition of acetyltransferases as part of the INHAT (inhibitor of histone acetyltransferases) complex. Plays a role in E4F1-mediated transcriptional repression. -
Tissue specificity
Expressed in all tissues tested. Highly expressed in kidney and skeletal muscle, moderate levels of expression in brain, placenta and pancreas, and weakly expressed in lung. Found in all regions of the brain examined (amygdala, caudate nucleus, corpus callosum, hippocampus and thalamus), with highest levels in amygdala. -
Sequence similarities
Belongs to the ANP32 family.
Contains 4 LRR (leucine-rich) repeats.
Contains 1 LRRCT domain. -
Post-translational
modificationsPhosphorylated on serine residues.
The N-terminus is blocked.
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. Cytoplasm. Endoplasmic reticulum. Translocates to the cytoplasm during the process of neuritogenesis (By similarity). Shuttles between nucleus and cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 8125 Human
- Entrez Gene: 11737 Mouse
- Entrez Gene: 25379 Rat
- Omim: 600832 Human
- SwissProt: P39687 Human
- SwissProt: O35381 Mouse
- SwissProt: P49911 Rat
- Unigene: 458747 Human
see all -
Alternative names
- acidic (leucine-rich) nuclear phosphoprotein 32 family, member A antibody
- Acidic leucine-rich nuclear phosphoprotein 32 family member A antibody
- Acidic nuclear phosphoprotein 32 family member A antibody
see all
Images
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Western blot - Anti-PHAP1 antibody (ab5991)All lanes : Anti-PHAP1 antibody (ab5991) at 0.5 µg/ml
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : ANP32A knockout HEK-293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : THP-1 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 33 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-PHAP1 antibody staining at 0.5 ug/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab5991 was shown to bind specifically to PHAP1. A band was observed at 33 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in ANP32A knockout cell line ab266148 (knockout cell lysate ab258303). To generate this image, wild-type and ANP32A knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Western blot - Anti-PHAP1 antibody (ab5991)All lanes : Anti-PHAP1 antibody (ab5991) at 1 µg/ml
Lane 1 : 293 cell lysate
Lane 2 : A431 cell lysate
Lane 3 : A549 cell lysate
Lane 4 : CaCo-2 cell lysate
Lane 5 : Daudi cell lysate
Lane 6 : HeLa cell lysate
Lane 7 : HepG2 cell lysate
Lane 8 : K562 cell lysate
Lane 9 : MCF-7 cell lysate
Lane 10 : Jurkat cell lysate
Lane 11 : SK-N-SH cell lysate
Lane 12 : THP-1 cell lysate
Lane 13 : NIH/3T3 cell lysate
Lane 14 : YB2/0 cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/10000 dilutionIncubated with the primary antibody for 1 hour at room temperature in 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHAP1 antibody (ab5991)ab5991 at 2µg/ml staining PHAP1 in mouse small intestine tissue by IHC
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Western blot - Anti-PHAP1 antibody (ab5991)All lanes : Anti-PHAP1 antibody (ab5991) at 1 µg/ml
Lane 1 : Mouse heart tissue lysate
Lane 2 : Mouse pancreas tissue lysate
Lane 3 : Mouse testis tissue lysate
Lane 4 : Mouse liver tissue lysate
Lane 5 : Mouse spleen tissue lysate
Lane 6 : Mouse kidney tissue lysate
Lane 7 : Mouse colon tissue lysate
Lane 8 : Mouse stomach tissue lysate
Lane 9 : Mouse brain tissue lysate
Lane 10 : Mouse skin tissue lysate
Lane 11 : Mouse skeletal muscle tissue lysate
Lane 12 : Mouse bladder tissue lysate
Lane 13 : Mouse lung tissue lysate
Lane 14 : K562 cell lysate
Lysates/proteins at 15 µg/ml per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/10000 dilutionIncubated with the primary antibody for 1 hour at room temperature in 5% NFDM/TBST.
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Western blot - Anti-PHAP1 antibody (ab5991)Western blot analysis of PHAP I expression in human Raji cell (A), mouse (B) and rat (C) testis tissue llysates with anti-PHAP I at 1 µg/ml.
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Immunocytochemistry/ Immunofluorescence - Anti-PHAP1 antibody (ab5991)Immunofluorescence of PHAP I in mouse small intestine cells using ab5991 at 20µg/ml.
Protocols
Datasheets and documents
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Datasheet download
References (4)
ab5991 has been referenced in 4 publications.
- Saavedra F et al. PP32 and SET/TAF-Iß proteins regulate the acetylation of newly synthesized histone H4. Nucleic Acids Res 45:11700-11710 (2017). PubMed: 28977641
- Wang S et al. The expression and distributions of ANP32A in the developing brain. Biomed Res Int 2015:207347 (2015). PubMed: 25866766
- Schafer ZT et al. Enhanced sensitivity to cytochrome c-induced apoptosis mediated by PHAPI in breast cancer cells. Cancer Res 66:2210-8 (2006). WB, Blocking ; Human . PubMed: 16489023
- Dirksen EH et al. Human lymphoblastoid proteome analysis reveals a role for the inhibitor of acetyltransferases complex in DNA double-strand break response. Cancer Res 66:1473-80 (2006). WB ; Human . PubMed: 16452203
