Anti-DUSP5 antibody [EPR19684]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DUSP5 antibody [EPR19684] (AB200708)

ab200708 staining DUSP5 in wild-type MEF cells serum starved and then treated for 2h with 20% FCS (top panel) and DUSP5 knockout MEF cells serum starved and then treated for 2h with 20% FCS (bottom panel). The cells were fixed with 100% methanol for 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab200708 at 1/100 dilution and anti-alpha Tubulin antibody ab7291 at 1/1000 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) secondary antibody and Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150120) at 2 μg/ml. Nuclear DNA was labelled in blue with DAPI.

• WB

Supplier Data

Western blot - Anti-DUSP5 antibody [EPR19684] (AB200708)

Blocking/Dilution buffer : 5% NFDM/TBST.

Human DUSP5 full length recombinant protein contain aa1-384 with a His-Tag®. This recombinant human full length protein was made in-house.

All lanes:

Western blot - Anti-DUSP5 antibody [EPR19684] (ab200708) at 1/5000 dilution

All lanes:

Human DUSP5 full length recombinant protein at 0.01 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 1/100000 dilution

Predicted band size: 42 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-DUSP5 antibody [EPR19684] (AB200708)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-DUSP5 antibody [EPR19684] (ab200708) at 1/1000 dilution

Lane 1:

HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Untreated MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

MCF7 (Human breast adenocarcinoma epithelial cell) treated with 100 ng/ml PMA for 48 hours whole cell lysate 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

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-DUSP5 antibody [EPR19684] (AB200708)

Blocking/Dilution buffer : 5% NFDM/TBST.

MEF Wt and DUSP5 KO whole cell lysates used on this blot are from MEFs serum starved and then treated for 2h with 20% FCS.

These lysates were kindly provided by Steve Keyes from Dundee University.

All lanes:

Western blot - Anti-DUSP5 antibody [EPR19684] (ab200708) at 1/1000 dilution

Lane 1:

MEF (Mouse embryonic fibroblast cell line) (WT) whole cell lysate at 20 µg

Lane 2:

MEF (Mouse embryonic fibroblast cell line) (DUSP5 KO) whole cell lysate 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 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-DUSP5 antibody [EPR19684] (AB200708)

Blocking/Dilution buffer : 5% NFDM/TBST.

Mouse DUSP5 full length recombinant protein contain aa1-384 with a His-Tag®. This recombinant mouse full length protein was made in-house.

All lanes:

Western blot - Anti-DUSP5 antibody [EPR19684] (ab200708) at 1/1000 dilution

All lanes:

Mouse DUSP5 full length recombinant protein at 0.01 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 1/100000 dilution

Predicted band size: 42 kDa

false

Exposure time: 4s