ICAM1 Mouse Monoclonal Antibody [F11-A4]

Western blot analysis of ICAM1 on different lysates with Mouse anti-ICAM1 antibody (M1505-7) at 1/1,000 dilution.

Lane 1: Ramos cell lysate
Lane 2: Raji cell lysate
Lane 3: HUVEC cell lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 58 kDa
Observed band size: 80 kDa

Exposure time: 2 minutes 30 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 (M1505-7) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

Western blot analysis of ICAM1 on different lysates with Mouse anti-ICAM1 antibody (M1505-7) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-ICAM1 KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 58 kDa
Observed band size: 80 kDa

Exposure time: 15 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 (M1505-7) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of Raji cells labeling ICAM1 with Mouse anti-ICAM1 antibody (M1505-7) 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 Mouse anti-ICAM1 antibody (M1505-7) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of Raji cells labeling ICAM1.

Cells were fixed and permeabilized. Then stained with the primary antibody (M1505-7, 1/1,000) (red) compared with Mouse 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).