Anti-ZO1 tight junction protein antibody [BLR092G]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZO1 tight junction protein antibody [BLR092G] (AB276131)

Paraffin embedded human kidney tissue stained for ZO1 tight junction protein using ab276131 at 1/500 dilution in immunohistochemical analysis. Secondary antibody used was HRP-conjugated goat anti-rabbit IgG. Substrate : DAB.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZO1 tight junction protein antibody [BLR092G] (AB276131)

Paraffin embedded human kidney tissue stained for ZO1 tight junction protein using ab276131 at 1/500 dilution in immunohistochemical analysis. Secondary antibody was used HRP-conjugated goat antirabbit IgG, with counterstaining seen in blue (DAPI).

• ICC

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Immunocytochemistry - Anti-ZO1 tight junction protein antibody [BLR092G] (AB276131)

A549 cells stained for ZO1 tight junction protein using ab276131 at 1/500 dilution in immunocytochemical analysis. Substrate : DAB

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZO1 tight junction protein antibody [BLR092G] (AB276131)

Paraffin embedded human prostate carcinoma tissue stained for ZO1 tight junction protein using ab276131 at 1/500 dilution in immunohistochemical analysis. Secondary antibody used was HRP-conjugated goat anti-rabbit IgG. Substrate : DAB.

• IP

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Immunoprecipitation - Anti-ZO1 tight junction protein antibody [BLR092G] (AB276131)

Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from A-549 cells was prepared using NETN lysis buffer. ab276131 was used for IP at 12 μl per reaction (Lane 1). TJP1/ZO-1 was also immunoprecipitated by rabbit anti-TJP1/ZO-1 antibody. For blotting immunoprecipitated ab276131 was used at 1/1000 dilution. Contol IgG shown in Lane 2.

Chemiluminescence with an exposure time of 30 seconds

All lanes:

Immunoprecipitation - Anti-ZO1 tight junction protein antibody [BLR092G] (ab276131)

Predicted band size: 195 kDa

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• WB

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Western blot - Anti-ZO1 tight junction protein antibody [BLR092G] (AB276131)

Chemiluminescence with an exposure time of 10 seconds.

Lower Panel : Rabbit antiCOPB2 antibody.

All lanes:

Western blot - Anti-ZO1 tight junction protein antibody [BLR092G] (ab276131) at 1/1000 dilution

Lane 1:

A549 whole cell lysate at 50 µg

Lane 2:

K562 whole cell lysate at 50 µg

Lane 3:

HEK-293T whole cell lysate at 50 µg

Lane 4:

MCF7 whole cell lysate at 50 µg

Lane 5:

Jurkat whole cell lysate at 50 µg

Lane 6:

HeLa whole cell lysate at 50 µg

Lane 7:

LNCaP whole cell lysate at 50 µg

Lane 8:

RKO whole cell lysate at 50 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG

Predicted band size: 195 kDa

false

• WB

Lab

Western blot - Anti-ZO1 tight junction protein antibody [BLR092G] (AB276131)

Western blot : Anti-TJP1 antibody [BLR092G] (ab276131) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab276131 was shown to bind specifically to TJP1. A band was observed at 195 kDa in wild-type HeLa cell lysates with no signal observed at this size in TJP1 knockout cell line. There is a close non-specific band at 190 kDa. We recommend running a low percentage gel to separate this from the band of interest. To generate this image, wild-type and TJP1 knockout HeLa 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-ZO1 tight junction protein antibody [BLR092G] (ab276131) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TJP1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-tjp1-zo1-tight-junction-protein-knockout-hela-cell-line-ab264897'>ab264897</a>) at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

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• WB

Supplier Data

Western blot - Anti-ZO1 tight junction protein antibody [BLR092G] (AB276131)

Chemiluminescence with an exposure time of 3 minutes.

Lower Panel : Rabbit anti-COPB2 antibody.

All lanes:

Western blot - Anti-ZO1 tight junction protein antibody [BLR092G] (ab276131) at 1/1000 dilution

Lane 1:

mIMCD-3 whole cell lysate at 50 µg

Lane 2:

RenCa whole cell lysate at 50 µg

Lane 3:

NIH/3T3 whole cell lysate at 50 µg

Lane 4:

TCMK-1 whole cell lysate at 50 µg

Lane 5:

C2C12 whole cell lysate at 50 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG

Predicted band size: 195 kDa

false