Recombinant 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
Related conjugates and formulations
Overview
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Product name
Anti-HRAS + KRAS antibody [EPR18713]
See all HRAS + KRAS primary antibodies -
Description
Rabbit monoclonal [EPR18713] to HRAS + KRAS -
Host species
Rabbit -
Specificity
Immunoprecipitation requires a mild lysis buffer to achieve an acceptable enrichment.
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Tested applications
Suitable for: ICC/IF, WB, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18713 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
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.
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WB | (5) |
1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 22, 21 kDa).
|
IP | (2) |
1/70.
Immunoprecipitation requires a mild lysis buffer to achieve an acceptable enrichment. |
Notes |
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ICC/IF
1/500. |
WB
1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 22, 21 kDa). |
IP
1/70. Immunoprecipitation requires a mild lysis buffer to achieve an acceptable enrichment. |
Target
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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
- Entrez Gene: 3265 Human
- Entrez Gene: 3845 Human
- Omim: 190020 Human
- Omim: 190070 Human
- SwissProt: P01112 Human
- SwissProt: P01116 Human
- SwissProt: P32883 Mouse
- SwissProt: Q61411 Mouse
see all -
Alternative names
- c-H-ras antibody
- c-K-ras antibody
- c-Ki-ras antibody
see all
Images
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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 secondBlocking/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.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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. -
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 analysis shows milder lysis buffer is required to get desirable enrichment.
Panel A: RIPA buffer
Panel B: IP lysis buffer (see protocol for details)
HRAS + KRAS was immunoprecipitated from Hepa1-6 (Mouse hepatoma epithelial cell) whole cell lysate 10µg with ab191595 at 1/30 dilution (2ug). Western blot was performed on the immunoprecipitate using ab191595 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1 & 4: Hepa1-6 (Mouse hepatoma epithelial cell) whole cell lysate 10µg
Lane 2 & 5: ab191595 IP in Hepa1-6 whole cell lysate
Lane 3 & 6: Rabbit monoclonal IgG (ab172730) instead of ab191595 in Hepa1-6 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
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