PACT (PKR activating protein) / PRKRA Recombinant Rabbit Monoclonal Antibody [PSH02-14]
cat.: HA721773
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, IF-Tissue, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH02-14 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 34 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human PACT aa 1-313 (O75569). |
| Positive control: | K-562cell lysate, HeLa cell lysate, MCF7 cell lysate, C2C12 cell lysate, PC-12 cell lysate, mouse testis tissue lysate, rat testis tissue lysate, human testis tissue, 293T cell lysate, K-562 cell lysate, Jurkat cell lysate, HepG2 cell lysate, Hela cell lysate, HL-60 cell lysate, mouse liver tissue lysate, rat skeletal muscle tissue lysate, rat brain tissue lysate, rat brain tissue, rat hippocampus tissue, 293T, PC-12. |
| Subcellular location: | Cytoplasm, perinuclear region Cytoplasm |
| Recommended Dilutions:
WB IHC-P IF-Cell IF-Tissue FC |
1:1,000-1:2,000 1:1,000~1:5,000 1:100 1: 200 1:1,000 |
| Uniprot #: | SwissProt: O75569 Human | Q9WTX2 Mouse | Q4V8C7 Rat |
| Alternative names: | DYT16 HSD14 Interferon inducible double stranded RNA dependent protein kinase activator A interferon-inducible double stranded RNA-dependent activator Interferon-inducible double stranded RNA-dependent protein kinase activator A PACT PKR associated protein X PKR associating protein X PKR-associated protein X PKR-associating protein X PRKRA PRKRA_HUMAN Protein activator of the interferon induced protein kinase Protein activator of the interferon-induced protein kinase Protein kinase Protein kinase interferon inducible double stranded RNA dependent activator RAX |
Images
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Fig1:
Western blot analysis of PACT (PKR activating protein) / PRKRA on different lysates with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: K-562cell lysate Lane 2: HeLa cell lysate Lane 3: MCF7 cell lysate Lane 4: C2C12 cell lysate Lane 5: PC-12 cell lysate Lane 6: Mouse testis tissue lysate Lane 7: Rat testis tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 34 kDa Observed band size: 34 kDa Exposure time: 30 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721773) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721773) at 1/1,000 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. |
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Fig3:
Western blot analysis of PACT (PKR activating protein) / PRKRA on different lysates with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/1,000 dilution. Lane 1: 293T cell lysate Lane 2: K-562 cell lysate Lane 3: Jurkat cell lysate Lane 4: HepG2 cell lysate Lane 5: MCF7 cell lysate Lane 6: Hela cell lysate Lane 7: HL-60 cell lysate Lane 8: C2C12 cell lysate Lane 9: PC-12 cell lysate Lane 10: Mouse testis tissue lysate Lane 11: Mouse liver tissue lysate Lane 12: Rat testis tissue lysate Lane 13: Rat skeletal muscle tissue lysate Lane 14: Rat brain tissue lysate Cell lysates: 20ug/lane, tissue lysates: 40ug/lane. Predicted band size: 34.4 kDa Observed band size: 34 kDa Exposure time: 1 minutes 2 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721773) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721773) at 1/5,000 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721773) at 1/5,000 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. |
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Fig6:
Immunocytochemistry analysis of 293T cells labeling PACT (PKR activating protein) / PRKRA with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig7:
Immunofluorescence analysis of paraffin-embedded rat brain tissue labeling PACT (PKR activating protein) / PRKRA with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721773, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig8:
Immunocytochemistry analysis of PC-12 cells labeling PACT (PKR activating protein) / PRKRA with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig9:
Immunofluorescence analysis of paraffin-embedded rat hippocampus tissue labeling PACT (PKR activating protein) / PRKRA with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721773, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig10:
Flow cytometric analysis of 293T cells labeling PACT (PKR activating protein) / PRKRA. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721773, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig11:
Western blot analysis of PACT (PKR activating protein) / PRKRA on different lysates with Rabbit anti-PACT (PKR activating protein) / PRKRA antibody (HA721773) at 1/2,000 dilution. Lane 1: HeLa-si NT cell lysate Lane 2: HeLa-si PRKRA cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 34 kDa Observed band size: 34 kDa Exposure time: 16 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721773) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature. |
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