HA721035

PIAS1 Recombinant Rabbit Monoclonal Antibody [JE30-36]

cat.: HA721035

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	JE30-36

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 68 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human PIAS1 aa 600-651.
Positive control:	MCF-7 cell lysate, HL-60 cell lysate, rat testis tissue lysate, rat testis tissue, human tonsil tissue, human thyroid tissue, human colon carcinoma tissue, mouse hippocampus tissue, mouse brain tissue, NCI-H441.
Subcellular location:	Nucleus, Nucleus speckle, Cytoplasm; Nucleus speckle, PML body, Nucleus; Nucleus speckle, PML body.
Recommended Dilutions: WB IF-Cell IHC-P	1:500 1:50 1:100-1:400
Uniprot #:	SwissProt: O75925 Human \| O88907 Mouse \| G3V9T0 Rat
Alternative names:	AR interacting protein DDXBP1 DEAD/H (Asp-Glu-Ala-Asp/His) box binding protein 1 DEAD/H box binding protein 1 DEAD/H box-binding protein 1 E3 SUMO-protein ligase PIAS1 GBP Gu binding protein Gu-binding protein GU/RH-II Pias1 PIAS1_HUMAN Protein inhibitor of activated STAT protein 1 Protein inhibitor of activated STAT, 1 RNA helicase II binding protein RNA helicase II-binding protein Zinc finger, MIZ-type containing 3 ZMIZ3

Images

	Fig1: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-PIAS1 antibody (HA721035) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721035) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig2: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-PIAS1 antibody (HA721035) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721035) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig3: Immunohistochemical analysis of paraffin-embedded human thyroid tissue with Rabbit anti-PIAS1 antibody (HA721035) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721035) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig4: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-PIAS1 antibody (HA721035) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721035) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-PIAS1 antibody (HA721035) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721035) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-PIAS1 antibody (HA721035) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721035) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: Immunocytochemistry analysis of NCI-H441 cells labeling PIAS1 with Rabbit anti-PIAS1 antibody (HA721035) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PIAS1 antibody (HA721035) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) were used as the secondary antibody at 1/1,000 dilution.

Fig8: Western blot analysis of PIAS1 on different lysates with Rabbit anti-PIAS1 antibody (HA721035) at 1/500 dilution.

Lane 1: MCF7 (Human breast cancer cells) cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 3 minutes; ECL: K1802

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA721035, 1/500 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 68.0 kDa
Observed band size: 75 kDa

Fig9: Western blot analysis of PIAS1 on different lysates with Rabbit anti-PIAS1 antibody (HA721035) at 1/500 dilution.

Lane 1: J774A.1(mouse mononuclear macrophages) cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 3 minutes; ECL: K1802

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA721035, 1/500 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 68.0 kDa
Observed band size: 75 kDa

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.