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Anti-PIAS1+PIAS2+PIAS3 Rabbit Monoclonal Antibody

Figure 1. Western blot analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human HepG2 whole cell lysates,
Lane 2: human Hacat whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human A431 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 8: mouse lung tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PIAS1+PIAS2+PIAS3 antigen affinity purified monoclonal antibody (Catalog # M01707) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PIAS1+PIAS2+PIAS3 at approximately 76 kDa. The expected band size for PIAS1+PIAS2+PIAS3 is at 76 kDa.
Figure 2. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of human colon adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 3. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of human colon adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 4. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 5. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 6. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of human thyroid papillary carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 7. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of human thyroid papillary carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 8. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of human urothelial carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 9. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of human urothelial carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 10. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of non-small cell lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 11. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of non-small cell lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 12. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of rat testis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 13. IHC analysis of PIAS1+PIAS2+PIAS3 using anti-PIAS1+PIAS2+PIAS3 antibody (M01707).
PIAS1+PIAS2+PIAS3 was detected in a paraffin-embedded section of rat testis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PIAS1+PIAS2+PIAS3 Antibody (M01707) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

SPECIFICATION
CITATIONS
PROTOCOLS
BACKGROUND

Product Information
Application Applications Legend: WB=Western Blot IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin-embedded Sections) IHC-F=Immunohistochemistry (Frozen Sections) IF=Immunofluorescence FC=Flow Cytopmetry IC=Immunochemistry ICC=Immunocytochemistry E=ELISA IP=Immunoprecipitation DB=Dot Blot CHIP=Chromatin Immunoprecipitation FA=Fluorescence Assay IEM=Immuno electron microscopy EIA=Enzyme Immunoassay	WB, IHC, IF, ICC, IP, FC
Primary Accession	O75925/O75928
Host	Rabbit
Isotype	Rabbit IgG
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Liquid
Description	Anti-PIAS1+PIAS2+PIAS3 Rabbit Monoclonal Antibody . Tested in WB, IHC, ICC/IF, IP, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.

Additional Information
Calculated MW	71836 MW KDa
Application Details	WB 1:500-1:2000 IHC 1:50-1:200 ICC/IF 1:50-1:200 IP 1:50 FC 1:50
Subcellular Localization	Nucleus speckle. Nucleus, PML body. Interaction with CSRP2 may induce a partial redistribution along the cytoskeleton.
Tissue Specificity	Expressed in numerous tissues with highest level in testis..
Contents	Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clone Names	Clone: ABFA-16
Immunogen	A synthesized peptide derived from human PIAS1+PIAS2+PIAS3
Purification	Affinity-chromatography
Storage	Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

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Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

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