Recombinant
RabMAb

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

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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

Overview

  • 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

    Suitable for: ICC/IF, WB, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant full length protein within Human HRAS + KRAS aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P01116-2

  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab191595 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/500.
WB 1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 22, 21 kDa).
IP 1/70.

Target

  • Relevance

    KRAS: Ras proteins bind GDP/GTP and possess intrinsic GTPase activity. Plays an important role in the regulation of cell proliferation (PubMed:23698361, PubMed:22711838). Enzyme regulation: Alternates between an inactive form bound to GDP and an active form bound to GTP. Activated by a guanine nucleotide-exchange factor (GEF) and inactivated by a GTPase-activating protein (GAP). Interaction with SOS1 promotes exchange of bound GDP by GTP. HRAS: Ras proteins bind GDP/GTP and possess intrinsic GTPase activity. Enzyme regulation: Alternates between an inactive form bound to GDP and an active form bound to GTP. Activated by a guanine nucleotide-exchange factor (GEF) and inactivated by a GTPase-activating protein (GAP).

  • Database links

    see all

  • Alternative names

    • c-H-ras antibody
    • c-K-ras antibody
    • c-Ki-ras antibody
    • GTPase HRas antibody
    • GTPase KRas antibody
    • H-Ras-1 antibody
    • Ha-Ras antibody
    • HRAS antibody
    • HRAS1 antibody
    • K-Ras 2 antibody
    • Ki-Ras antibody
    • KRAS antibody
    • KRAS2 antibody
    • p21ras antibody
    • RASK2 antibody
    • Transforming protein p21 antibody
    see all

Images

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

References

Publishing research using ab191595? Please let us know so that we can cite the reference in this datasheet.

ab191595 has been referenced in 1 publication.

  • Thu PM  et al. Phellodendrine chloride suppresses proliferation of KRAS mutated pancreatic cancer cells through inhibition of nutrients uptake via macropinocytosis. Eur J Pharmacol 850:23-34 (2019). PubMed: 30716311

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