NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody [PSH09-98]

☑ Relative expression (RE)

Western blot analysis of NG2/CSPG4 on different lysates with Rabbit anti-NG2/CSPG4 antibody (HA723167) at 1/10,000 dilution.

Lane 1: A375 cell lysate
Lane 2: SK-MEL-28 cell lysate
Lane 3: MCF7 cell lysate (negative)
Lane 4: HeLa cell lysate
Lane 5: PANC-1 cell lysate
Lane 6: MDA-MB-231 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 251 kDa
Observed band size: 330 kDa

Exposure time: Lane 1-2: 6 seconds; Lane 3-6: 59 seconds; ECL: K1801;

3-8% 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 (HA723167) at 1/10,000 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.

☑ Relative expression (RE)

Immunocytochemistry analysis of A375 (positive) and SK-Br-3 (negative) labeling NG2/CSPG4 with Rabbit anti-NG2/CSPG4 antibody (HA723167) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-NG2/CSPG4 antibody (HA723167) 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 (HA601187, 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.

☑ Relative expression (RE)

Flow cytometric analysis of SK-Br-3 (left, negative) and A375 (right, positive) cells labeling NG2/CSPG4.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723167, 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).