Anti-Integrin beta 4 抗体 [EPR17517]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Integrin beta 4 with ab182120 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Membrane and cytoplasm staining on Human colon epithelium is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling Integrin beta 4 with ab182120 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Membrane and cytoplasm staining on Human cervical carcinoma is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling Integrin beta 4 with ab182120 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cell junction staining on mouse pancreatic acinus is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• WB

Unknown

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (ab182120) at 1/5000 dilution

All lanes:

SW480 (Human colon adenocarcinoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 202 kDa

Observed band size: 210 kDa

false

Exposure time: 5s

• WB

Lab

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Lane 1 : Wild-type HAP1 whole cell lysate (40 μg)
Lane 2 : ITGB4 knockout HAP1 whole cell lysate (40 μg)

Lanes 1 - 2 : Merged signal (red and green). Green - ab182120 observed at 240 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab182120 was shown to specifically recognize ITGB4 in wild-type HAP1 cells alond with additional cross-reactive bands. No band was observed when ITGB4 knockout samples were used. Wild-type and ITGB4 knockout samples were subjected to SDS-PAGE. ab182120 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/30,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (ab182120)

Predicted band size: 202 kDa

false

• WB

Lab

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Western blot : Anti-ITGB4 antibody [EPR17517] (ab182120) 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, ab182120 was shown to bind specifically to ITGB4. A band was observed at 200 kDa in wild-type A549 cell lysates with no signal observed at this size in ITGB4 knockout cell line. To generate this image, wild-type and ITGB4 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-Integrin beta 4 antibody [EPR17517] (ab182120) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 40 µg

Lane 2:

ITGB4 knockout A549 cell lysate at 40 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: 200 kDa

false

• WB

Supplier Data

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (ab182120) at 1/20000 dilution

All lanes:

Human skin lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 202 kDa

Observed band size: 210 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (ab182120) at 1/1000 dilution

All lanes:

Human pancreas lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 202 kDa

Observed band size: 210 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (ab182120) at 1/1000 dilution

All lanes:

BxPC-3 (Human pancreas adenocarcinoma cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 202 kDa

Observed band size: 210 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (ab182120) at 1/1000 dilution

Lane 1:

Mouse pancreas lysate at 10 µg

Lane 2:

Mouse ovary lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 202 kDa

Observed band size: 210 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (ab182120) at 1/1000 dilution

Lane 1:

Mouse spleen lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 202 kDa

Observed band size: 210 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Integrin beta 4 antibody [EPR17517] (ab182120) at 1/20000 dilution

All lanes:

Mouse skin lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 202 kDa

Observed band size: 210 kDa

false

Exposure time: 15s