Name: Anti-Ras (mutated Q61R) antibody [SP174]
SKU: ab227658
Rating: 4 (4 reviews)

Anti-Ras antibody (Mutated-Q61R) [SP174] ab227658 is a rabbit monoclonal antibody that is used in Ras western blotting, IHC and flow cytometry. Suitable for human samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Validated on the Leica BOND™ RX automated IHC staining platform for IHC
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras (mutated Q61R) antibody [SP174] (AB227658), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.6
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	Flow Cyt (Intra)	WB	IHC-P
Human	Tested	Tested	Tested
Transfected cell line - Human	Not recommended	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/2000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell line - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell line - Human	Dilution info 1/400 - 1/1000	Notes -
Species Human	Dilution info 1/400 - 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell line - Human	Dilution info 1/100	Notes Perform heat mediated antigen retrieval with EDTA buffer pH 8.0 before commencing with IHC staining protocol. This antibody may not be able to detect endogenous NRAS Q61K in patient tissue samples by IHC-P based on a collaborator's data.
Species Human	Dilution info 1/100	Notes Perform heat mediated antigen retrieval with EDTA buffer pH 8.0 before commencing with IHC staining protocol. This antibody may not be able to detect endogenous NRAS Q61K in patient tissue samples by IHC-P based on a collaborator's data.

Associated Products

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Target data

See full target information NRAS mutated Q61R

Function

Ras proteins bind GDP/GTP and possess intrinsic GTPase activity.

Alternative names

HRAS1, NRAS, GTPase NRas, Transforming protein N-Ras

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.6
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: SP174
Purification technique: Affinity purification Protein A/G
Specificity: This antibody has been found to react with different mutations we tested. We have not tested all possible mutations at Q61.
Concentration
Purification notes: Purified from TCS by protein A/G.

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The Ras protein also known by its alternate names such as KRAS NRAS and HRAS functions as a small GTPase which is a type of enzyme that binds to guanosine triphosphate (GTP) and can switch to an inactive form when it hydrolyzes GTP to GDP. Ras proteins are encoded by genes located in different chromosomes and typically have a molecular weight of about 21 kDa. These proteins are expressed in various tissues throughout the body where they play key roles in cellular signaling.

Biological function summary

Ras proteins are pivotal in regulating cell proliferation differentiation and survival. They are often part of larger protein complexes that facilitate signal transduction across cell membranes. These proteins function as binary molecular switches toggling between active (GTP-bound) and inactive (GDP-bound) states. Mutations in Ras proteins such as NRAS Q61R can lead the protein to assume permanently active conformations disrupting normal cellular signaling processes and contributing to oncogenesis.

Pathways

Ras proteins play significant roles in the MAPK/ERK and PI3K/AKT signaling pathways. Their activation leads to a cascade of phosphorylation events that ultimately regulate gene expression. Within these pathways Ras proteins interact with various molecules including RAF kinases and the PI3K protein further illustrating their complex roles in signal propagation. These pathways control many cellular processes including growth differentiation and survival illustrating how Ras proteins integrate multiple signals to modulate cellular outcomes.

Associated diseases and disorders

Ras proteins are frequently implicated in various cancers notably pancreatic and colorectal cancers. Mutated forms of Ras such as KRAS G12V drive oncogenesis by promoting uncontrolled cell growth and division. In cancer pathology Ras often interacts with tumor suppressor proteins influencing the disease progression. Targeting Ras-mediated signaling pathways using approaches like pan-Ras inhibitors or chemiluminescence ELISAs represents an ongoing area of therapeutic research aimed at combating Ras-driven malignancies.

Product promise

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Full details and terms and conditions can be found here:
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10 product images

This image is courtesy of an anonymous collaborator.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
Immunohistochemical analysis of paraffin-embedded human primary malignant melanoma tissue (mutation status: NRAS Q61R) labeling NRAS with ab227658 at 1/100 dilution for 30 min at RT. The tissue was counterstained with Hematoxylin. Heat mediated antigen retrieval was achieved by using EDTA buffer pH 9.0 before IHC staining.
This image is courtesy of an anonymous collaborator.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
Immunohistochemical analysis of paraffin-embedded human primary malignant melanoma tissue (mutation status: NRAS Q61R) labeling NRAS with ab227658 at 1/100 dilution for 30 min at RT. The tissue was counterstained with Hematoxylin. Heat mediated antigen retrieval was achieved by using EDTA buffer pH 9.0 before IHC staining.
This image is courtesy of an anonymous collaborator.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
Immunohistochemical analysis of paraffin-embedded human primary malignant melanoma tissue (mutation status: NRAS Q61R) labeling NRAS with ab227658 at 1/100 dilution for 30 min at RT. The tissue was counterstained with Hematoxylin. Heat mediated antigen retrieval was achieved by using EDTA buffer pH 9.0 before IHC staining.
This image is courtesy of an anonymous collaborator.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
Immunohistochemical analysis of paraffin-embedded human metastatic malignant melanoma tissue (mutation status: NRAS Q61R) labeling NRAS with ab227658 at 1/100 dilution for 30 min at RT. The tissue was counterstained with Hematoxylin. Heat mediated antigen retrieval was achieved by using EDTA buffer pH 9.0 before IHC staining.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
Immunohistochemical analysis of paraffin-embedded human metastatic malignant melanoma tissue (mutation status: NRAS Q61K) labeling NRAS with ab227658 at 1/100 dilution for 30 min at RT. The tissue was counterstained with Hematoxylin. Heat mediated antigen retrieval was achieved by using EDTA buffer pH 9.0 before IHC staining.
Flow Cytometry (Intracellular) - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
Flow Cytometry analysis of SK-MEL-2( Human skin malignant melanoma) cells labeling NRAS with purifiedab227658 at 1:200 dilution (0.8 μg/ml) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor© 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Black. Unlabeled control - Unlabelled cells / blue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
IHC image of NRAS (mutated Q61R) staining in a section of formalin-fixed paraffin-embedded human skin melanoma* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH9, epitope retrieval solution 2) for 20mins. The section was then incubated with ab227658, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
All lanes: Western blot - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658) at 1/400 dilution
All lanes: SK-MEL-2 cell lysate
Predicted band size: 21 kDa
Western blot - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM /TBST
This antibody has been found to react with different mutations we tested. We haven’t tested all possible mutations at Q61.
All lanes: Western blot - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658) at 1/1000 dilution
Lane 1: HEK-293T (Human embryonic kidney epithelial cell) transfected with an empty vector (vector control), containing a His-tag, whole cell lysate at 20 µg
Lane 2: HEK-293T (Human embryonic kidney epithelial cell) transfected with His-tagged NRAS (mutated Q61R) whole cell lysate at 20 µg
Lane 3: HEK-293T (Human embryonic kidney epithelial cell) transfected with His-tagged NRAS (mutated Q61K) whole cell lysate at 20 µg
Lane 4: HEK-293T (Human embryonic kidney epithelial cell) transfected with His-tagged NRAS (mutated Q61H) whole cell lysate at 20 µg
Lane 5: HEK-293T (Human embryonic kidney epithelial cell) transfected with His-tagged NRAS (mutated Q61L) whole cell lysate at 20 µg
Lane 6: HEK-293T (Human embryonic kidney epithelial cell) transfected with His-tagged NRAS whole cell lysate at 20 µg
Lane 7: HEK-293T (Human embryonic kidney epithelial cell) transfected with His-tagged HRAS (mutated Q61R) whole cell lysate at 20 µg
Lane 8: HEK-293T (Human embryonic kidney epithelial cell) transfected with His-tagged KRAS (mutated Q61R) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 21 kDa
Observed band size: 25 kDa
Exposure time: 5s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras (mutated Q61R) antibody [SP174] (ab227658)
Immunohistochemistry analysis of paraffin-embedded (A) HEK-293T transfected with NRAS (Q61R) expression vector, (B) HEK-293T transfected with NRAS (Q61K) expression vector, (C) HEK-293T transfected with NRAS (Q61H) expression vector, (D) HEK-293T transfected with NRAS (Q61L) expression vector, (E) HEK-293T transfected with NRAS expression vector, (F) HEK-293T transfected with HRAS (Q61R) expression vector, (G) HEK-293T transfected with KRAS(Q61R) and (H) HEK-293T transfected empty vector, sections labelling NRAS (mutated Q61R) with ab227658 at 1/100 dilution. The section was incubated with ab227658 for 10 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.
Positive staining on HEK-293T transfected with NRAS (Q61R) expression vector (Image A), HEK-293T transfected with NRAS (Q61K) expression vector (Image B), HEK-293T transfected with HRAS (Q61R) expression vector (Image F) and HEK-293T transfected with KRAS (Q61R) expression vector (Image G); Nearly no staining on HEK-293T transfected with NRAS (Q61H) expression vector (Image C), HEK-293T transfected with NRAS (Q61L) expression vector (Image D), HEK-293T transfected with NRAS expression vector (Image E) and HEK-293T transfected empty vector (Image H). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.