Anti-MICA + MICB antibody [EPR22031] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MICA + MICB antibody [EPR22031] - BSA and Azide free (AB241513)

Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling MICA with ab224702 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on epithelium of human stomach. (PMID : 28334733) is observed. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab224702).

• WB

Lab

Western blot - Anti-MICA + MICB antibody [EPR22031] - BSA and Azide free (AB241513)

This data was developed using the same antibody clone in a different buffer formulation (abAB224702).

Western blot : Anti-MICA antibody [EPR22031] (ab224702) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab224702 was shown to bind specifically to MICA. A band was observed at 43 kDa in wild-type A549 cell lysates with no signal observed at this size in MICA knockout cell line. To generate this image, wild-type and MICA 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-MICA + MICB antibody [EPR22031] (<a href='/en-us/products/primary-antibodies/mica-micb-antibody-epr22031-ab224702'>ab224702</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

MICA knockout A549 cell lysate at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

Ramos cell lysate at 20 µg

Lane 5:

MICA protein <a href='/en-us/products/proteins-peptides/recombinant-human-mica-protein-active-ab182709'>ab182709</a> cell lysate at 0.2 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: 43 kDa

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