Recombinant Anti-NRAS antibody [EPR25186-24] (ab300431)
![Western blot - Anti-NRAS antibody [EPR25186-24] (AB300431)](https://www.abcam.com/ps/products/300/ab300431/Images/AB300431-487884-rabbit-anti-nras-epr25186-24-wstern-blot-hek-293t-ko.jpg "Western blot - Anti-NRAS antibody [EPR25186-24] (AB300431)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25186-24] to NRAS
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-NRAS antibody [EPR25186-24]
See all NRAS primary antibodies -
Description
Rabbit monoclonal [EPR25186-24] to NRAS -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: Flow Cyt (Intra),ICC/IF or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Whole cell lysates: Wild-type HEK-293T, MCF7 (human breast adenocarcinoma epithelial cell), A375 (human malignant melanoma epithelial cell) , C6 (rat glial tumor glial cell), NIH/3T3 (mouse embryonic fibroblast) . Tissue lysates: Rat and mouse spleen. His-tagged human NRAS recombinant protein. IP: MCF7 (human breast adenocarcinoma epithelial cell).
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General notes
ab300431 does not react in IHC-P with human, mouse, and rat; ICC with human, and mouse, and intrecellular flow cytometry with mouse.
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: 7.20
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
EPR25186-24 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab300431 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 |
|---|---|---|
| WB |
1/1000.
|
|
| IP |
1/30.
|
| Notes |
|---|
|
WB
1/1000. |
|
IP
1/30. |
Target
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Function
Ras proteins bind GDP/GTP and possess intrinsic GTPase activity. -
Involvement in disease
Defects in NRAS are a cause of juvenile myelomonocytic leukemia (JMML) [MIM:607785]. JMML is a pediatric myelodysplastic syndrome that constitutes approximately 30% of childhood cases of myelodysplastic syndrome (MDS) and 2% of leukemia.
Defects in NRAS are the cause of Noonan syndrome type 6 (NS6) [MIM:613224]. A syndrome characterized by facial dysmorphic features such as hypertelorism, a downward eyeslant and low-set posteriorly rotated ears. Other features can include short stature, a short neck with webbing or redundancy of skin, cardiac anomalies, deafness, motor delay and variable intellectual deficits. -
Sequence similarities
Belongs to the small GTPase superfamily. Ras family. -
Post-translational
modificationsPalmitoylated by the ZDHHC9-GOLGA7 complex. A continuous cycle of de- and re-palmitoylation regulates rapid exchange between plasma membrane and Golgi. -
Cellular localization
Cell membrane. Golgi apparatus membrane. Shuttles between the plasma membrane and the Golgi apparatus. - Information by UniProt
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Database links
- Entrez Gene: 4893 Human
- Entrez Gene: 18176 Mouse
- Entrez Gene: 24605 Rat
- Omim: 164790 Human
- SwissProt: P01111 Human
- SwissProt: P08556 Mouse
- SwissProt: Q04970 Rat
- Unigene: 486502 Human
see all -
Alternative names
- ALPS4 antibody
- AV095280 antibody
- GTPase NRas antibody
see all
Images
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Western blot - Anti-NRAS antibody [EPR25186-24] (AB300431)All lanes :
Lane 1 : Wild-type HEK-293T (ab255449) whole cell lysate
Lane 2 : NRAS knockout HEK-293T (ab266684) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 21 kDa why is the actual band size different from the predicted?
Exposure time: 180 secondsBlocking and dilution buffer and concentration: 5% NFDM/TBST.
In Western blot, ab300431 was shown to bind specifically to NRAS. One band was observed at 21 kDa in wild-type HEK-293T (ab255449) cell lysates with no signal observed at this size in NRAS knockout cell lysates (ab258542).
This blot was developed using a high sensitivity ECL substrate. -
![Western blot - Anti-NRAS antibody [EPR25186-24] (AB300431)](https://www.abcam.com/ps/products/300/ab300431/Images/AB300431-487472-anti-nras-antibody-epr2518624-western-blot-human-kras-recombinant-protein-10-ng.jpg)
Western blot - Anti-NRAS antibody [EPR25186-24] (AB300431)All lanes : Anti-NRAS antibody [EPR25186-24] (ab300431) at 1/1000 dilution
Lane 1 : Human KRAS recombinant protein, 10 ng
Lane 2 : His-tagged human HRAS recombinant protein, 10 ng
Lane 3 : His-tagged human NRAS recombinant protein, 10 ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 21 kDa why is the actual band size different from the predicted?
Exposure time: 37 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
This antibody does not cross-react with human KRAS or HRAS.
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![Western blot - Anti-NRAS antibody [EPR25186-24] (AB300431)](https://www.abcam.com/ps/products/300/ab300431/Images/AB300431-487475-anti-nras-antibody-epr2518624-western-blot-mcf7-human-breast-adenocarcinoma-epithelial-cell-whole-cell-lysate-40-ug.jpg)
Western blot - Anti-NRAS antibody [EPR25186-24] (AB300431)All lanes : Anti-NRAS antibody [EPR25186-24] (ab300431) at 1/1000 dilution
Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : A375 (human malignant melanoma epithelial cell) whole cell lysate
Lane 3 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 4 : Rat spleen tissue lysate
Lane 5 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 6 : Mouse spleen tissue lysate
Lysates/proteins at 40 µg per lane.
Secondary
Lanes 1-3 & 5-6 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051))
Observed band size: 21 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a high sensitivity ECL substrate.
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![Immunoprecipitation - Anti-NRAS antibody [EPR25186-24] (AB300431)](https://www.abcam.com/ps/products/300/ab300431/Images/AB300431-2-anti-nras-antibody-epr2518624-immunoprecipitation-mcf7.jpg)
Immunoprecipitation - Anti-NRAS antibody [EPR25186-24] (AB300431)NRAS was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab300431 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300431 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: MCF7 whole cell lysate 10 μg
Lane 2: ab300431 IP in MCF7 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300431 in MCF7 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds
Observed MW (kDa): 21.
This blot was developed using a high sensitivity ECL substrate.
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![Anti-NRAS antibody [EPR25186-24] (AB300431)](https://www.abcam.com/ps/products/300/ab300431/Images/AB300431-1-Benefits-of-Recombinant-antibodies.jpg)
Anti-NRAS antibody [EPR25186-24] (AB300431)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab300431 has not yet been referenced specifically in any publications.