Anti-MKK7 antibody [EP1455Y]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MKK7 antibody [EP1455Y] (AB52618)

Intracellular Flow Cytometry analysis ofMCF7 (Human breast adenocarcinoma epithelial cell) cells labeling MKK7 with ab52618at 1/100 dilution (1μg) (Red). Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody. Cells were fixed with4% paraformaldehyde. Rabbit monoclonal IgG (ab172730) was used as isotype control (Black). Unlabelled control : Cells without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MKK7 antibody [EP1455Y] (AB52618)

ab52618 at 1/50 dilution staining human baldder tissue; paraffin embedded.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-MKK7 antibody [EP1455Y] (AB52618)

Lanes 1, 5 and 9 : Wild-type HAP1 cell lysate (20 μg)
Lanes 2, 6 and 10 : MEK7 knockout HAP1 cell lysate (20 μg)
Lanes 3, 7 and 11 : HeLa cell lysate (20 μg)
Lanes 4, 8 and 12 : A431 cell lysate (20 μg)
Lanes 1, 2, 3 and 4 : Green signal from target - ab52618 observed at 47 kDa
Lanes 5, 6, 7 and 8 : Red signal from loading control - ab8245 observed at 37 kDa
Lanes 9, 10, 11 and 12 : Merged (red and green) signal
ab52618 was shown to specifically react with MEK7 when MEK7 knockout samples were used. Wild-type and MEK7 knockout samples were subjected to SDS-PAGE. ab52618 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-MKK7 antibody [EP1455Y] (ab52618)

Predicted band size: 47 kDa

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• WB

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Western blot - Anti-MKK7 antibody [EP1455Y] (AB52618)

All lanes:

Western blot - Anti-MKK7 antibody [EP1455Y] (ab52618) at 1/10000 dilution

All lanes:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP antibody at 1/2000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

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• WB

Supplier Data

Western blot - Anti-MKK7 antibody [EP1455Y] (AB52618)

Dilution and blocking buffer : 5% NFDM/TBST.

Low expression control : mouse and rat skeletal muscle.

In Western blot, anti-Vinculin antibody (ab129002) loading control staining at 1/10000 dilution.

All lanes:

Western blot - Anti-MKK7 antibody [EP1455Y] (ab52618) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate (Low expression control) at 20 µg

Lane 4:

Rat testis tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat skeletal muscle tissue lysate (Low expression control) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 47 kDa

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Exposure time: 15s