Anti-Ras antibody [EPR18713-13]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Ras antibody [EPR18713-13] (AB206969)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling RAS with ab206969 at 1/200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm and nuclear staining on HeLa cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab206969 at 1/200 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Ras antibody [EPR18713-13] (AB206969)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling RASwith ab206969 at 1/250 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Ras antibody [EPR18713-13] (AB206969)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling RAS with ab206969 at 1/200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm and nuclear staining on NIH/3T3 cells line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab206969 at 1/200 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-Ras antibody [EPR18713-13] (AB206969)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 1 second; Lane 2 and 3 : 5 seconds.

Recombinant Human RAS full length protein contains aa1-189 with an N-terminal His-Tag®. Recombinant Human HRAS full length protein contains aa1-189 with an N-terminal His-Tag®. Recombinant Human NRAS full length protein contains aa1-189 with an N-terminal His-Tag®. All three recombinant proteins were made in house.

All lanes:

Western blot - Anti-Ras antibody [EPR18713-13] (ab206969) at 1/5000 dilution

Lane 1:

Recombinant Human KRAS full length protein at 0.01 µg

Lane 2:

Recombinant Human HRAS full length protein at 0.01 µg

Lane 3:

Recombinant Human NRAS full length protein at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• WB

Supplier Data

Western blot - Anti-Ras antibody [EPR18713-13] (AB206969)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. This antibody cross-reacts with human HRAS/NRAS/KRAS. In Western blot, anti-Ras antibody (ab206969) staining at 1/1000 dilution. Exposure time : 26 seconds.

All lanes:

Western blot - Anti-RAS antibody [18/Ras] (<a href='/ko/products/primary-antibodies/ras-antibody-18-ras-ab307738'>ab307738</a>) at 1/1000 dilution

Lane 1:

His-tagged human HRAS recombinant protein at 10 ng

Lane 2:

His-tagged human NRAS recombinant protein at 10 ng

Lane 3:

His-tagged human KRAS recombinant protein at 10 ng

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 21 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Ras antibody [EPR18713-13] (AB206969)

Blocking/Dilution buffer : 5% NFDM/TBST.

Lane 1:

Western blot - Anti-Ras antibody [EPR18713-13] (ab206969) at 1/2000 dilution

Lane 2:

Western blot - Anti-Ras antibody [EPR18713-13] (ab206969) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Ras antibody [EPR18713-13] (AB206969)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Ras antibody [EPR18713-13] (ab206969) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

A431 (Human epidermoid carcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Ras antibody [EPR18713-13] (AB206969)

Blocking/Dilution buffer : 5% NFDM/TBST.

Lane 1:

Western blot - Anti-Ras antibody [EPR18713-13] (ab206969) at 1/2000 dilution

Lane 2:

Western blot - Anti-Ras antibody [EPR18713-13] (ab206969) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Ras antibody [EPR18713-13] (AB206969)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Ras antibody [EPR18713-13] (ab206969) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Rat brain lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

Exposure time: 3min