CD276 Recombinant Rabbit Monoclonal Antibody [PSH02-46]
cat.: HA721823
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH02-46 |
| 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: | Full length recombinant protein within human CD276. |
| Positive control: | HeLa cell lysate, MCF7 cell lysate, HEK-293 cell lysate, LoVo cell lysate, U-2 OS cell lysate, LNCaP cell lysate, SH-SY5Y cell lysate, THP-1 cell lysate, HCT 116 cell lysate, THP-1, MCF7. |
| Subcellular location: | Membrane. |
| Recommended Dilutions:
WB IF-Cell FC |
1:2,000-5,000 1:100 1:1,000 |
| 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 (HA721823) at 1/2,000 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: 42 seconds; ECL: K1802; 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 (HA721823) 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 (HA721823) at 1/5,000 dilution. Lane 1: HAP1-parental cell lysate Lane 2: HAP1-CD276 KD cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 57 kDa Observed band size: 100 kDa Exposure time: 180 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 (HA721823) at 1/5,000 dilution was used in K1803 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:
Immunocytochemistry analysis of THP-1 cells labeling CD276 with Rabbit anti-CD276 antibody (HA721823) 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-CD276 antibody (HA721823) 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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Fig4:
Immunocytochemistry analysis of MCF7 cells labeling CD276 with Rabbit anti-CD276 antibody (HA721823) at 1/100 dilution. Cells were fixed in 100% precooled methanol 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-CD276 antibody (HA721823) 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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Fig5:
Flow cytometric analysis of THP-1 cells labeling CD276. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA721823, 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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