Anti-CD18 antibody [MEM-148]

• Flow Cyt

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Flow Cytometry - Anti-CD18 antibody [MEM-148] (AB8220)

Human peripheral blood lymphocytes stained with ab8220 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID : 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab8220, 0.1μg/1x10⁶ cells) for 30 min at 4°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD18 antibody [MEM-148] (AB8220)

Flow cytometry surface staining pattern of human peripheral whole blood stained using ab8220.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD18 antibody [MEM-148] (AB8220)

Separation of human monocytes (red-filled) from CD18 negative blood debris (black-dashed) in flow cytometry analysis (surface staining) of human peripheral whole blood stained using ab8220.

• WB

Supplier Data

Western blot - Anti-CD18 antibody [MEM-148] (AB8220)

The reactivity of MEM-148 antibody was assessed by comparing binding signals in a panel of various human cell samples. Western blotting analysis was performed on RIPA buffer extracts of THP-1, Kg1a, U937, HPB-ALL, Raji, Jurkat, leukocytes, and HeLa cells, mixed and heated (100°C, 5 min) with reducing (2-mercaptoethanol) SDS-loading buffer. Samples were resolved using 10% SDS-PAGE gel.

Nitrocellulose membrane blot was probed with ab8220 (2 µg/ml), followed by IRDye 800CW Goat-anti-Mouse IgG (green). For multiplex fluorescent Western blot detection, mouse anti-GAPDH monoclonal antibody FF26A conjugated with DyLight 680 (0.1 µg/ml) was used as the loading control (red).

CD18 was detected at ~90-100 kDa in the respective cell lines.

All lanes:

Western blot - Anti-CD18 antibody [MEM-148] (ab8220) at 2 µg/mL

Lane 1:

THP-1

Lane 2:

Kg1a

Lane 3:

U937

Lane 4:

HPB-ALL

Lane 5:

Raji

Lane 6:

Jurkat

Lane 7:

Leukocytes

Lane 8:

HeLa

Secondary

All lanes:

IRDye 800CW Goat-anti-Mouse IgG

Observed band size: 90-100 kDa

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