Recombinant
RabMAb

Recombinant Anti-KRAS+HRAS+NRAS antibody [EPR18713-13] (ab206969)

Overview

  • Product name

    Anti-KRAS+HRAS+NRAS antibody [EPR18713-13]
    See all KRAS+HRAS+NRAS primary antibodies
  • Description

    Rabbit monoclonal [EPR18713-13] to KRAS+HRAS+NRAS
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • WB: Recombinant human full length KRAS, NRAS and HRAS proteins; Human fetal brain and fetal kidney lysates; HeLa, A431, C6 and NIH/3T3 whole cell lysates; Mouse and rat brain lysates. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: HeLa cells. IP: A549 whole cell lysate.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • 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

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18713-13
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab206969 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 1/200.
Flow Cyt 1/250.
IP 1/70.
WB 1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).

Target

Images

  • All lanes : Anti-KRAS+HRAS+NRAS antibody [EPR18713-13] (ab206969) at 1/5000 dilution

    Lane 1 : Recombinant Human KRAS full length protein
    Lane 2 : Recombinant Human HRAS full length protein
    Lane 3 : Recombinant 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 1/100000 dilution

    Predicted band size: 21 kDa
    Observed band size: 21 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

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

    Recombinant Human KRAS 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.

  • Lane 1 : Anti-KRAS+HRAS+NRAS antibody [EPR18713-13] (ab206969) at 1/2000 dilution
    Lane 2 : Anti-KRAS+HRAS+NRAS antibody [EPR18713-13] (ab206969) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    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


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-KRAS+HRAS+NRAS antibody [EPR18713-13] (ab206969) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 21 kDa
    Observed band size: 21 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Lane 1 : Anti-KRAS+HRAS+NRAS antibody [EPR18713-13] (ab206969) at 1/2000 dilution
    Lane 2 : Anti-KRAS+HRAS+NRAS antibody [EPR18713-13] (ab206969) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 21 kDa
    Observed band size: 21 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-KRAS+HRAS+NRAS antibody [EPR18713-13] (ab206969) 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 1/100000 dilution

    Predicted band size: 21 kDa
    Observed band size: 21 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling KRAS+HRAS+NRAS 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling KRAS+HRAS+NRAS 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.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling KRAS+HRAS+NRAS with 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.

  • KRAS+HRAS+NRAS was immunoprecipitated from 1mg of A549 (Human lung carcinoma) whole cell lysate with ab206969 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab206969 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: A549 whole cell lysate 10ug (Input).

    Lane 2: ab206969 IP in A549 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab206969 in A549 whole cell lysate.

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

    Exposure time: 10 seconds.

References

This product has been referenced in:

  • Wang L  et al. TIPE-2 suppresses growth and aggressiveness of hepatocellular carcinoma cells through downregulation of the phosphoinositide 3-kinase/AKT signaling pathway. Mol Med Rep 17:7017-7026 (2018). Read more (PubMed: 29568863) »
See 1 Publication for this product

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