CD276 Recombinant Rabbit Monoclonal Antibody [PD00-36]
cat.: HA721245
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | IHC-P, FC, WB |
| Clonality: | Monoclonal |
| Clone number: | PD00-36 |
| 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: 57 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within C terminal human CD276. |
| Positive control: | Human lung squamous cell carcinoma tissue, human lung adenocarcinoma tissue, human prostate carcinoma tissue, HEK-293, THP-1. |
| Subcellular location: | Membrane. |
| Recommended Dilutions:
IHC-P FC WB |
1:5,000 1:500-1:1,000 1:2,000-1:5,000 |
| Uniprot #: | SwissProt: Q5ZPR3 Human |
| Alternative names: | 4Ig B7 H3 4Ig-B7-H3 AU016588 B7 H3 B7 homolog 3 B7-H3 B7H3 B7RP-2 CD_antigen=CD276 CD276 CD276 antigen CD276 molecule CD276_HUMAN Costimulatory molecule Flags: Precursor PSEC0249 UNQ309/PRO352 |
Images
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Fig1:
Western blot analysis of CD276 on different lysates with Rabbit anti-CD276 antibody (HA721245) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: MCF7 cell lysate Lane 3: HEK-293 cell lysate Lane 4: LoVo cell lysate Lane 5: U-2 OS cell lysate Lane 6: LNCaP cell lysate Lane 7: SH-SY5Y cell lysate Lane 8: THP-1 cell lysate Lane 9: HCT 116 cell lysate Lane 10: Raji cell lysate(negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 57 kDa Observed band size: 100 kDa Exposure time: 6 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 (HA721245) at 1/2,000 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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Fig2:
Western blot analysis of CD276 on different lysates with Rabbit anti-CD276 antibody (HA721245) at 1/5,000 dilution. Lane 1: HS-SY-II-si NT cell lysate Lane 2: HS-SY-II-si CD276 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 57 kDa Observed band size: 90 kDa Exposure time: 1 minute 2 seconds; ECL: K1801; 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 (HA721245) at 1/5,000 dilution was used in primary antibody dilution 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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Fig3:
Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma tissue with Rabbit anti-CD276 antibody (HA721245) 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 (HA721245) 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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Fig4:
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma tissue with Rabbit anti-CD276 antibody (HA721245) 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 (HA721245) 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 human prostate carcinoma tissue with Rabbit anti-CD276 antibody (HA721245) 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 (HA721245) 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:
Flow cytometric analysis of HEK-293 cells labeling CD276. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721245, 1ug/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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Fig7:
Flow cytometric analysis of THP-1 cells labeling CD276. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721245, 1ug/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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