Anti-LAIR1 antibody [EPR24745-12A]

• Flow Cyt

Lab

Flow Cytometry - Anti-LAIR1 antibody [EPR24745-12A] (AB315967)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab315967(right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab315967 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control(1x 10⁶ in 100 μl at 0.2 μg/ml (1/25)) for 30min on ice. The cells were simultaneously stained with CD14.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/2000 for 30min on ice.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable monocytes.

• Flow Cyt

Lab

Flow Cytometry - Anti-LAIR1 antibody [EPR24745-12A] (AB315967)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab315967 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab315967 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶ in 100 μl at 0.2 μg/ml (1/25)) for 30min on ice. The cells were simultaneously stained with CD19.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/2000 for 30min on ice.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable CD3(-) lymphocytes.

• WB

Lab

Western blot - Anti-LAIR1 antibody [EPR24745-12A] (AB315967)

Lanes 1 & 2 : Merged signal (red and green). Green – ab315967 observed at 40 kDa. Red - loading control, ab7291 observed at 50 kDa.

ab315967 was shown to specifically react with LAIR1. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. ab315967 and Anti-alpha Tubulin antibody [DM1A]- Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilutions, respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-LAIR1 antibody [EPR24745-12A] (ab315967) at 1/1000 dilution

Lane 1:

THP-1 cell lysate at 20 µg

Lane 2:

HeLa cell lysate

Secondary

Lanes 1 - 2:

Goat anti-Rabbit IgG H&L (IRDye® 800CW) at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L (IRDye® 680RD) at 1/20000 dilution

Observed band size: 40 kDa,50 kDa

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