Anti-DAP12 antibody

• WB

Lab

Western blot - Anti-DAP12 antibody (AB93846)

False colour image of Western blot : Anti-DAP12 antibody staining at 1 ug/ml, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab93846 was shown to bind specifically to DAP12. A band was observed at 12 kDa in wild-type THP-1 cell lysates with no signal observed at this size in TYROBP CRISPR-Cas9 edited cell line ab273726 (CRISPR-Cas9 edited cell lysate ab273774). The band observed in the CRISPR-Cas9 edited lysate lane below 12 kDa is likely to represent a truncated form of DAP12. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and TYROBP CRISPR-Cas9 edited THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % BSA in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 2 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-DAP12 antibody (ab93846) at 1 µg/mL

Lane 1:

Wild-type THP-1 cell lysate at 40 µg

Lane 2:

TYROBP CRISPR-Cas9 edited THP-1 cell lysate at 40 µg

Predicted band size: 12 kDa

Observed band size: 12 kDa

false

• WB

Lab

Western blot - Anti-DAP12 antibody (AB93846)

False colour image of Western blot : Anti-DAP12 antibody staining at 1 ug/ml, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab93846 was shown to bind specifically to DAP12. A band was observed at 12 kDa in wild-type THP-1 cell lysates with no signal observed at this size in TYROBP knockout cell line ab273726 (knockout cell lysate ab273774). The band observed in the knockout lysate lane below 12 kDa is likely to represent a truncated form of DAP12. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and TYROBP knockout THP-1 cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % BSA in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 2 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-DAP12 antibody (ab93846) at 1 µg/mL

Lane 1:

Wild-type THP-1 cell lysate at 40 µg

Lane 2:

TYROBP knockout THP-1 cell lysate at 40 µg

Lane 2:

Western blot - Human TYROBP (DAP12) knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-tyrobp-dap12-knockout-thp-1-cell-line-ab273726'>ab273726</a>)

Predicted band size: 12 kDa

Observed band size: 12 kDa

false

• WB

Unknown

Western blot - Anti-DAP12 antibody (AB93846)

The DAP12 protein sequence contains a 21-residue signal sequence at the N-terminus. Cleavage of this peptide may account for the doublet observed.

All lanes:

Western blot - Anti-DAP12 antibody (ab93846) at 1 µg/mL

All lanes:

Lung (Human) Tissue Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

Predicted band size: 12 kDa

Observed band size: 12 kDa,200 kDa

true

Exposure time: 12min

• WB

Lab

Western blot - Anti-DAP12 antibody (AB93846)

ab93846 was shown to react with TYROBP in wild-type THP-1 cells in Western blot with loss of signal observed in a TYROBP knockout cell line. Wild-type THP-1 and TYROBP knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab93846 overnight at 4 °C at a 1/1000  dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-DAP12 antibody (ab93846) at 1/1000 dilution

Lane 1:

Wild-type THP-1 lysate at 100 µg

Lane 2:

TYROBP knock-out THP-1 lysate at 100 µg

false