Anti-TREM2 antibody [EPR20243]

• IP

Supplier Data

Immunoprecipitation - Anti-TREM2 antibody [EPR20243] (AB209814)

TREM2 was immunoprecipitated from 0.35 mg of THP-1 (Human monocytic leukemia cell line) whole cell lysate with ab209814 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab209814 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : THP-1 whole cell lysate, 10 μg (Input).

Lane 2 : ab209814 IP in THP-1 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab209814 in THP-1 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-TREM2 antibody [EPR20243] (ab209814)

Predicted band size: 25 kDa

Observed band size: 25 kDa

false

• WB

Lab

Western blot - Anti-TREM2 antibody [EPR20243] (AB209814)

Lanes 1 - 3 : Merged signal (red and green). Green - ab209814 observed at 30 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab209814 was shown to react with TREM2 in wild-type THP-1 cells in Western blot with loss of signal observed in TREM2 knockout cell line ab269489 (TREM2 knockout cell lysate ab269652). Wild-type THP-1 and TREM2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab209814 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-TREM2 antibody [EPR20243] (ab209814) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human TREM2 knockout THP-1 cell lysate (<a href='/en-us/products/cell-lysates/human-trem2-knockout-thp-1-cell-lysate-ab269652'>ab269652</a>) at 20 µg

Lane 3:

HL-60 cell lysate at 20 µg

Predicted band size: 25 kDa

Observed band size: 30 kDa

false

• WB

Supplier Data

Western blot - Anti-TREM2 antibody [EPR20243] (AB209814)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-TREM2 antibody [EPR20243] (ab209814) at 1/1000 dilution

Lane 1:

Untreated THP-1 (Human monocytic leukemia cell line) whole cell lysate at 20 µg

Lane 2:

THP-1 whole cell lysate treated with 100ng/ml Phorbol-12-myristate-13-acetate (PMA) for 72 hours at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 25 kDa

Observed band size: 26 kDa

false

Exposure time: 4s