Anti-STRAD antibody [EPR15603]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-STRAD antibody [EPR15603] (AB192879)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling STRAD antibody (red) with purified ab192879 at a dilution of 1/200. Goat anti rabbit IgG (Alexa Fluor^® 488) was used as the secondary antibody at 1/2000. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-STRAD antibody [EPR15603] (AB192879)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% triton X-100 permeabilized A549 cells labeling STRAD with ab192879 at a 1/400 dilution, followed by Goat anti rabbit IgG(Alexa Fluor® 488) secondary antibody (ab150077) at 1/400 dilution. Nuclear counter stain : Dapi (blue).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STRAD antibody [EPR15603] (AB192879)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling STRAD with ab192879 at a 1/100 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG as a secondary antibody. Counter stained with Hematoxylin. Negative control also shown.

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STRAD antibody [EPR15603] (AB192879)

Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue labeling STRAD with ab192879 at a 1/100 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG as a secondary antibody. Counter stained with Hematoxylin. Negative control also shown.

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

• WB

Lab

Western blot - Anti-STRAD antibody [EPR15603] (AB192879)

Western blot : Anti-STRAD antibody [EPR15603] ab192879 staining at 1/5000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 48 kDa in Wild-type A549 cell lysates with no signal observed at this size in STRADA knockout A549 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

Lanes 1 - 5:

Western blot - Anti-STRAD antibody [EPR15603] (ab192879) at 1/5000 dilution

Lanes 1 - 5:

Western blot - Anti-STRAD antibody [EPR15603] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/strad-antibody-epr15603-bsa-and-azide-free-ab232340'>ab232340</a>) at 1/5000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

STRADA knockout A549 at 20 µg

Lane 3:

MCF7 at 20 µg

Lane 4:

U-87 MG at 20 µg

Lane 5:

HT-29 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 48 kDa

Observed band size: 48 kDa

false

• WB

Supplier Data

Western blot - Anti-STRAD antibody [EPR15603] (AB192879)

All lanes:

Western blot - Anti-STRAD antibody [EPR15603] (ab192879) at 1/5000 dilution

Lane 1:

293 cell lysate at 20 µg

Lane 2:

HepG2 cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Human fetal brain lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 48 kDa

false