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
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
HA721823_1.jpg 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.
HA721823_2.jpg Fig2: 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.
HA721823_3.jpg Fig3: 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.
HA721823_4.jpg Fig4: 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.