Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
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Species reactivity: | Human, Mouse, Rat |
Applications: | WB, IHC-P, IF-Cell, FC, IP |
Clonality: | Monoclonal |
Clone number: | PSH01-55 |
Form: | Liquid |
Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
Storage buffer: | PBS (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: 130 kDa |
Isotype: | IgG |
Immunogen: | Recombinant protein within Human PCB aa 450-1178. |
Positive control: | HepG2 cell lysate, MCF7 cell lysate, A549 cell lysate, Huh7 cell lysate, Hela cell lysate, NIH/3T3 cell lysate, rat brain tissue lysate, mouse liver tissue, mouse lymph node tissue, rat liver tissue, rat brain tissue, HepG2, PC-12. |
Subcellular location: | Mitochondrion matrix. |
Recommended Dilutions:
WB IHC-P IF-Cell FC IP |
1:1,000 1:200 1:250 1:1,000 1-2μg/sample |
Uniprot #: | SwissProt: P11498 Human | Q05920 Mouse | P52873 Rat |
Alternative names: | mitochondrial OTTHUMP00000235155 OTTHUMP00000235156 PC PCB Pcx PYC_HUMAN Pyruvate carboxylase Pyruvate carboxylase mitochondrial Pyruvic carboxylase |
Fig1:
Western blot analysis of PCB on different lysates with Rabbit anti-PCB antibody (HA721698) at 1/1,000 dilution. Lane 1: HepG2 cell lysate Lane 2: MCF7 cell lysate Lane 3: A549 cell lysate Lane 4: Huh7 cell lysate Lane 5: Hela cell lysate Lane 6: NIH/3T3 cell lysate Lane 7: Rat brain tissue lysate Lysates/proteins at 30 µg/Lane. Predicted band size: 130 kDa Observed band size: 130 kDa Exposure time: 24 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 (HA721698) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 mouse liver tissue with Rabbit anti-PCB antibody (HA721698) 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721698) 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 mouse lymph node tissue with Rabbit anti-PCB antibody (HA721698) 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721698) 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-PCB antibody (HA721698) 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721698) 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-PCB antibody (HA721698) 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721698) 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. |
Fig6:
Immunocytochemistry analysis of HepG2 cells labeling PCB with Rabbit anti-PCB antibody (HA721698) 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-PCB antibody (HA721698) 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:
Immunocytochemistry analysis of PC-12 cells labeling PCB with Rabbit anti-PCB antibody (HA721698) at 1/250 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-PCB antibody (HA721698) at 1/250 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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Fig8:
Flow cytometric analysis of PC-12 cells labeling PCB. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721698, 1/1,000) (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). |
Fig9:
PCB was immunoprecipitated in 0.2mg HeLa cell lysate with HA721698 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA721698 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: Rabbit IgG instead of HA721698 in HeLa cell lysate Lane 3: HA721698 IP in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 24 seconds |