Anti-MERTK antibody [Y323] - Low endotoxin, Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [Y323] - Low endotoxin, Azide free (AB176887)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lymphoma tissue labeling MERTK with unpurified ab52968 at a 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab52968).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [Y323] - Low endotoxin, Azide free (AB176887)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labeling MERTK with purified ab52968 at 1/500.

Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051 a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Counterstained with hematoxylin.

Negative control using PBS instead of primary antibody (Inset).

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab52968).

• IP

Unknown

Immunoprecipitation - Anti-MERTK antibody [Y323] - Low endotoxin, Azide free (AB176887)

Lane 1 : ab52968 (purified) at 1/20 immunoprecipitating MERTK in HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate.

Lane 2 - PBS.

For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking/Dilution buffer and concentration : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52968).

All lanes:

Immunoprecipitation - Anti-MERTK antibody [Y323] (<a href='/en-us/products/primary-antibodies/mertk-antibody-y323-ab52968'>ab52968</a>)

Predicted band size: 110 kDa

Observed band size: 140-180 kDa

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• WB

Lab

Western blot - Anti-MERTK antibody [Y323] - Low endotoxin, Azide free (AB176887)

This data was developed using the same antibody clone in a different buffer formulation (ab52968 ).

Western blot : Anti-MERTK antibody [Y323] (ab52968) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52968 was shown to bind specifically to MERTK. A band was observed at 160 kDa in wild-type A549 cell lysates with no signal observed at this size in MERTK knockout cell line. To generate this image, wild-type and MERTK knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-MERTK antibody [Y323] (<a href='/en-us/products/primary-antibodies/mertk-antibody-y323-ab52968'>ab52968</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

MERTK knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human MERTK knockout A549 cell line (<a href='/en-us/products/cell-lines/human-mertk-knockout-a549-cell-line-ab301074'>ab301074</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 160 kDa

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