- Datasheet
- Specific References (8)
- Protocols
Overview
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Product nameAnti-PIAS1 antibody
See all PIAS1 primary antibodies -
DescriptionRabbit polyclonal to PIAS1
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Host speciesRabbit
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Tested applicationsSuitable for: ICC/IF, WB, IP, IHC-P, ChIPmore details
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Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat, Chicken, Dog, Xenopus laevis
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Immunogen
Synthetic peptide derived from within residues 500 - 600 of Human PIAS1.
(Peptide available as ab32218.)
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Positive control
- This antibody gave a positive signal when tested against Recombinant PIAS1 (Human) Protein. It also gave a positive signal in human testis tissue lysate.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
Applications
Our Abpromise guarantee covers the use of ab32219 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/IF | Use a concentration of 1 µg/ml. | |
| WB | 1/500 - 1/2000. Detects a band of approximately 81 kDa (predicted molecular weight: 72 kDa). | |
| IP | Use at an assay dependent concentration. | |
| IHC-P | 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| ChIP | Use at an assay dependent concentration. |
Target
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FunctionFunctions as an E3-type small ubiquitin-like modifier (SUMO) ligase, stabilizing the interaction between UBE2I and the substrate, and as a SUMO-tethering factor. Plays a crucial role as a transcriptional coregulation in various cellular pathways, including the STAT pathway, the p53 pathway and the steroid hormone signaling pathway. In vitro, binds A/T-rich DNA. The effects of this transcriptional coregulation, transactivation or silencing, may vary depending upon the biological context. Together with PRMT1, may repress STAT1 transcriptional activity, in the late phase of interferon gamma (IFN-gamma) signaling.
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Tissue specificityExpressed in numerous tissues with highest level in testis.
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PathwayProtein modification; protein sumoylation.
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Sequence similaritiesBelongs to the PIAS family.
Contains 1 PINIT domain.
Contains 1 SAP domain.
Contains 1 SP-RING-type zinc finger. -
DomainThe LXXLL motif is a transcriptional coregulator signature.
The SP-RING-type domain is required for promoting EKLF sumoylation. -
Post-translational
modificationsSumoylated.
Dimethylated by PRMT1 at Arg-303 in the late phase of interferon gamma (IFN-gamma) signaling, leading to preferential interaction with STAT1 and thus resulting in release of STAT1 from its target gene. -
Cellular localizationNucleus speckle. Interaction with CSRP2 may induce a partial redistribution along the cytoskeleton.
- Information by UniProt
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Database links
- Entrez Gene: 427514 Chicken
- Entrez Gene: 8554 Human
- Entrez Gene: 56469 Mouse
- Entrez Gene: 300772 Rat
- Omim: 603566 Human
- SwissProt: O75925 Human
- SwissProt: O88907 Mouse
- Unigene: 162458 Human
see all -
Alternative names
- AR interacting protein antibody
- DDXBP1 antibody
- DEAD/H (Asp-Glu-Ala-Asp/His) box binding protein 1 antibody
see all
Images
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Western blot - Anti-PIAS1 antibody (ab32219)This image is courtesy of an Abreview submitted by Min young KimAll lanes : Anti-PIAS1 antibody (ab32219) at 1/2500 dilution (Incubation for 1 hour at 25°C.)
Lane 1 : 293T cell lysate
Lane 2 : MEL cell lysate
Lane 3 : K562 cell lysate
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : Donkey Anti-Rabbit IgG H&L (HRP) (ab6802) at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 71 kDa (why is the actual band size different from the predicted?)
Exposure time: 1 minute
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Western blot - Anti-PIAS1 antibody (ab32219)All lanes : Anti-PIAS1 antibody (ab32219) at 1/1000 dilution
Lane 1 : HeLa whole cell lysate at 25 µg
Lane 2 : HeLa whole cell extract with transfected PIAS1 at 25 µg
Lane 3 : HeLa nuclear extract at 25 µg
Lane 4 : T47D nuclear extract
at 25 µg
Lane 5 : HeLa whole cell lysate immunoprecipitated with ab32219
Lane 6 : HeLa whole cell extract with transfected PIAS1 immunoprecipitated with ab32219
Lane 7 : HeLa nuclear extract immunoprecipitated with ab32219
Lane 8 : T47D nuclear extract immunoprecipitated with ab32219
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 81 kDa (why is the actual band size different from the predicted?)
Additional bands at: 110 kDa, 79 kDa (possible cleavage fragment), 79 kDa (possible cross reactivity). We are unsure as to the identity of these extra bands.
ab32219 was used in western blot analysis using HeLa whole cell lysate, HeLa nuclear lysate and T47D nuclear lysate. It was unable to detect endogenous PIAS1 in these lysates. It was able to detect PIAS1 in cells that are overexpressing the protein, running at a molecular weight of approximately 81kDa, suggesting that the amount of endogenous PIAS1 in these cells is very low. When ab32219 was used to immunoprecipitate the PIAS1 from these extracts, the protein could then be detected by western blot in the overexpressing HeLa extract as well as the T47D nuclear extract.For immunoprecipitations, 25ug of extract was used in the IP, and 10% of the sample was run on the gel. -
Immunocytochemistry/ Immunofluorescence - Anti-PIAS1 antibody (ab32219)ICC/IF image of ab32219 stained MEF1 cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab32219, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PIAS1 antibody (ab32219)Image courtesy of Human Protein Atlas
ab32219 staining PIAS1 in human stomach. Paraffin embedded human stomach tissue was incubated with ab32219 (1/200 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab32219 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.
Further results for this antibody can be found at www.proteinatlas.org
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Western blot - Anti-PIAS1 antibody (ab32219)Anti-PIAS1 antibody (ab32219) at 1 µg/ml + PIAS1 - Recombinant Protein at 0.1 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 100 kDa (why is the actual band size different from the predicted?)
Exposure time: 2 minutesab32219 recognizes the full length tagged recombinant protein which has an expected molecular weight of 98 kDa. -
Western blot - Anti-PIAS1 antibody (ab32219)Anti-PIAS1 antibody (ab32219) at 1 µg/ml + Human testis tissue lysate - total protein (ab30257) at 10 µg
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 72 kDa
Additional bands at: 180 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutes
Protocols
References
This product has been referenced in:
- Ibata M et al. Leukemogenic kinase FIP1L1-PDGFRA and a small ubiquitin-like modifier E3 ligase, PIAS1, form a positive cross-talk through their enzymatic activities. Cancer Sci 108:200-207 (2017). Read more (PubMed: 27960034) »
- Wang R et al. Reversible regulation of ORC2 SUMOylation by PIAS4 and SENP2. Oncotarget 8:70142-70155 (2017). Read more (PubMed: 29050267) »
Customer reviews and Q&As
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