Anti-VISTA antibody [BLR035F]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VISTA antibody [BLR035F] (AB243891)

Formalin-fixed, paraffin-embedded human lung carcinoma tissue stained for VISTA using ab243891 at 1/250 dilution in immunohistochemical analysis. A HRP-conjugated goat anti-rabbit antibody was used as the secondary. DAB staining.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VISTA antibody [BLR035F] (AB243891)

Immunohistochemistry analysis of formalin-fixed, paraffin-embedded human ovarian carcinoma tissue labeling VISTA with ab243891 at a 1/250 dilution, followed by HRP-conjugated Goat Anti-Rabit IgG secondary antibody. Substrate : Opal. Counterstain : DAPI (blue).

• ICC

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Immunocytochemistry - Anti-VISTA antibody [BLR035F] (AB243891)

Formalin-fixed, paraffin-embedded NCI H226 cells labeling VISTA using ab243891 at 1/250 dilution in ICC analysis. A HRP-conjugated goat-anti rabbit IgG was used as the secondary. DAB staining.

• IP

Unknown

Immunoprecipitation - Anti-VISTA antibody [BLR035F] (AB243891)

VISTA was immunoprecipitated from 1.0 mg Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab243891 at 20 µl per reaction. Western blot was performed on the immunoprecipitate using ab243891 at 1/1000 dilution. Lane 1 : ab243891 IP in Jurkat whole cell lysate.

Lane 2 : Contol IgG in Jurkat whole cell lysate.

Detection : Chemiluminescence with an exposure time of 30 seconds.

All lanes:

Immunoprecipitation - Anti-VISTA antibody [BLR035F] (ab243891)

Predicted band size: 34 kDa

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• WB

Lab

Western blot - Anti-VISTA antibody [BLR035F] (AB243891)

Western blot : Anti-VSIR antibody [BLR035F] (ab243891) 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, ab243891 was shown to bind specifically to VSIR. A band was observed at 50-60 kDa in wild-type A549 cell lysates with no signal observed at this size in VSIR knockout cell line. To generate this image, wild-type and VSIR 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-VISTA antibody [BLR035F] (ab243891) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

VSIR knockout A549 cell lysate at 20 µg

Lane 3:

DU 145 cell lysate at 20 µg

Lane 4:

MCF7 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

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• WB

Unknown

Western blot - Anti-VISTA antibody [BLR035F] (AB243891)

Western blot analysis using ab243891 at 1/1000 dilution.

Lane 1 : Jurkat whole cell lysate (50 μg).

Lane 2 : MOLT-4 whole cell lysate (50 μg).

Lane 3 : MJ whole cell lysate (50 μg).

Lane 4 : RPMI 8226 whole cell lysate (50 μg).

Lane 5 : HeLa whole cell lysate (50 μg).

Lane 6 : KG-1 whole cell lysate (50 μg).

Lane 7 : HEK-293T whole cell lysate (50 μg).

Lane 8 : SUP-M2 whole cell lysate (50 μg).

A HRP-conjugated goat anti-rabbit IgG antibody was used as the secondary. Detection : chemiluminescence with an exposure time of 30 seconds.

All lanes:

Western blot - Anti-VISTA antibody [BLR035F] (ab243891)

Predicted band size: 34 kDa

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