Antibodies, Proteins, Kits and Reagents for Life Science

294 835 ₸ Product size

100 µl 294 835 ₸ 10 µl 117 264 ₸

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Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18713] to HRAS + KRAS
Suitable for: ICC/IF, WB, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-HRAS + KRAS antibody [EPR18713]
See all HRAS + KRAS primary antibodies
Description

Rabbit monoclonal [EPR18713] to HRAS + KRAS
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Mouse Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human KRAS and HRAS full length proteins; Human fetal kidney and fetal brain lysates; Mouse and Rat brain lysates; A549, C6, A431 and RAW 264.7 whole cell lysates. ICC/iF: HeLa and NIH/3T3 cells. IP: A549 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18713
Isotype

IgG
Research areas

Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

All lanes : Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/10000 dilution

Lane 1 : Human KRAS full length protein
Lane 2 : Human HRAS full length protein
Lane 3 : Human NRAS full length protein

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa
why is the actual band size different from the predicted?

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.
Human KRAS full length protein contains aa1-189 with His-Tag® at N-Terminus; Human HRAS full length protein contains aa1-189 with His-Tag® at N-Terminus; Human NRAS full length protein contains aa1-189 with His-Tag® at N-Terminus. These are all made in-house.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/2000 dilution + Human fetal kidney lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/5000 dilution + Human fetal brain lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

All lanes : Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/2000 dilution + A549 (Human lung carcinoma) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

All lanes : Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/5000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysate
Lane 3 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling KRAS + HRAS with ab191595 at 1/500 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: ab191595 at 1/500 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.
Immunocytochemistry/ Immunofluorescence - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling KRAS + HRAS with ab191595 at 1/500 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: ab191595 at 1/500 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.
Immunoprecipitation - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

KRAS was immunoprecipitated from 1mg of A549 (Human lung carcinoma) whole cell lysate with ab191595 at 1/70 dilution.
Western blot was performed from the immunoprecipitate using ab191595 at 1/1000 dilution.
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: A549 whole cell lysate 10ug (Input).
Lane 2: ab191595 IP in A549 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191595 in A549 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

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