- Tested Species Reactivity:
- Predicted Species Reactivity:
- Cow, Dog, Horse, Human
- Product Format:
- Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
- Reconstitution and Storage:
- For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
- Replacement Item:
- This antibody may replace item sc-14016 from Santa Cruz Biotechnology.
- The immunogen is a synthetic peptide directed towards the middle region of human PIAS1
- Affinity Purified
- Predicted Homology Based on Immunogen Sequence:
- Cow: 79%; Dog: 93%; Horse: 79%; Human: 100%
- Complete computational species homology data:
- Anti-PIAS1 (ARP39232_P050)
- Peptide Sequence:
- Synthetic peptide located within the following region: LPTTNGSSSGSNSSLVSSNSLRESHSHTVTNRSSTDTASIFGIIPDIISL
- Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
- Blocking Peptide:
- For anti-PIAS1 (ARP39232_P050) antibody is Catalog # AAP39232 (Previous Catalog # AAPY00844)
- Printable datasheet for anti-PIAS1 (ARP39232_P050) antibody
- Sample Type Confirmation:
There is BioGPS gene expression data showing that PIAS1 is expressed in COLO205
- Target Reference:
- Helbig,K.J., (2008) Antiviral Res. 77 (3), 169-176
- Gene Symbol:
- Official Gene Full Name:
- Protein inhibitor of activated STAT, 1
- Alias Symbols:
- DDXBP1, GBP, GU/RH-II, MGC141878, MGC141879, ZMIZ3
- NCBI Gene Id:
- Protein Name:
- E3 SUMO-protein ligase PIAS1
- Description of Target:
- PIAS1 is a member of the mammalian PIAS [protein inhibitor of activated STAT-1 (signal transducer and activator of transcription-1)] family. This member contains a putative zinc-binding motif and a highly acidic region. It inhibits STAT1-mediated gene act
- Swissprot Id:
- Protein Accession #:
- Nucleotide Accession #:
- Protein Size (# AA):
- Molecular Weight:
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express PIAS1.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express PIAS1.
- Protein Interactions:
- STAT1; TRIM63; TRIM55; CEP250; PRDM1; SUMO1; L3MBTL2; CBS; SHFM1; SUMO2; SUMO3; SOX2; CHUK; SPOP; YWHAZ; UBC; SKIL; SGTA; ATXN1; SMAD7; SMAD4; SMAD1; HTT; PRPF40A; PPP1CC; BARD1; PAXIP1; NR1H2; PPARGC1A; NR1H3; PPARGC1B; RAD54L2; ZMYND11; UBD; PLAG1; MYB;
Application: Western blotting
Species + Tissue/Cell type: Lane 1: Untreated human U251 lysate Lane 2: IL-1beta treated human U251 lysate
Primary antibody dilution: 1:500
Secondary antibody: Anti-rabbit-HRP
Secondary antibody dilution: 1:500
|How do Aviva's reagents play a role in your experimental goals?||Regulation of the expression of PIAS1 in glial cells|
|How would you rate this antibody on a scale from 1-5 (5=best) and why?||5 - not work properly|
|Would you use this antibody in future experiments?||Probably|
|Have you used another antibody which has worked in your application?||No|
|Do you believe the information about the reagent on Aviva's website is correct?||Yes|
|If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?||May be|
|How did you store the antibody after re-suspension?||-20C|
|Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):||Human; Glioblastoma cells|
|How many different experimental trials were conducted using the antibody sample?||2|
|How was this sample prepared?||Cell cultures in 60 mm dishes were scraped into lysis buffer (PBS with a cocktail of protease inhibitors (Sigma) plus 1% Triton X-100), incubated for 30 min. at 4C with constant rotation, and centrifuged at 10,000 rpm for 10 min.|
|Primary antibody dilution and incubation time:||1:500|
|Secondary antibody used and dilution and incubation time:||1:500|
|Please include your detailed WB Procedure/Protocol here:||Western blotting.
1. Cell cultures in 60 mm dishes were scraped into lysis buffer (PBS with a cocktail of protease inhibitors (Sigma) plus 1% Triton X-100), incubated for 30 min. at 4C with constant rotation, and centrifuged at 10,000 rpm for 10 min. The protein concentration was measured with Bio-Rad Protein assay (#500-0006).
2. Forty micrograms of protein was separated by electrophoresis in 10% sodium dodecyl sulfate-polyacrylamide gel and then transferred to polyvinylidene difluoride membrane (Bio-Rad#162-0177) at 60V for 3h in cold room (4C).
3. The blots were blocked in 5% nonfat milk (Bio-Rad #170-6404) prepared in PBS + 0.1% Tween-20 for 1 h at room temperature (RT) with constant shaking.
4. Then the membranes were incubated with Primary antibodies o/n (~18h) at 4C with constant rotation.
5. After washing (shaking 3 times for 5 min each in PBS+0.1% Tween-20) the Secondary antibodies (horseradish peroxidase-conjugated anti-rabbit IgG (Thermo Scientific/Pierce#1858415)) were applied at dilution 1:500 - 1:1,000 for 1 h at RT with constant rotation.
6. After washing (3 times for 5 min each) the signals were detected with SuperSignal West Pico (or Femto) Chemiluminescent Substrate (Thermo Scientific/Pierce #34077 or #34095).
7. All blots were reprobed for b-actin to control the protein loading. At first, the membranes were stripped with Restore PLUS Western Blot Stripping Buffer (Thermo Scientific#46430), then blocked in 5% milk (see step 3), and then incubated with anti-Actin Ab (1:500) o/n at 4C, washed, and incubated with horseradish peroxidase-conjugated anti-rabbit IgG (1:500) followed washing and ECL development.
We used CL-XPosure Film (Thermo Scientific#34090).