Anti-Ras antibody [EPR23474-20] - BSA and Azide free
- BOND RX™ Validated
- Advanced Validation
- RabMAb
- Recombinant
- What is this?
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Rabbit Recombinant Monoclonal RASH antibody. Carrier free. Suitable for mIHC, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Rat, Recombinant full length protein - Human samples.
View Alternative Names
HRAS1, HRAS, GTPase HRas, H-Ras-1, Ha-Ras, Transforming protein p21, c-H-ras, p21ras
- IHC-P
Unknown
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Ras with ab275875 at 1/2000 (0.338 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human colon. The section was incubated with ab275875 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
- Flow Cyt (Intra)
Unknown
Flow Cytometry (Intracellular) - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling Ras with ab275875 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
A Goat anti rabbit IgG (Alexa Fluor®488, ab150077) at 1/2000 dilution was used as the secondary antibody.
- IP
Unknown
Immunoprecipitation - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Ras was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab275875 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275875 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2 : ab275875 IP in HeLa whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab275875 in HeLa whole cell lysate
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 110 seconds.
All lanes:
Immunoprecipitation - Anti-Ras antibody [EPR23474-20] (<a href='/en-us/products/primary-antibodies/ras-antibody-epr23474-20-ab275875'>ab275875</a>)
Predicted band size: 21 kDa
Observed band size: 21 kDa
false
- IP
Unknown
Immunoprecipitation - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Ras was immunoprecipitated from 0.35 mg A431 (human epidermoid carcinoma epithelial cell) whole cell lysate 10 with ab275875 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275875 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1 : A431 (human epidermoid carcinoma epithelial cell) whole cell lysate 10 ug
Lane 2 : ab275875 IP in A431 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab275875 in A431 whole cell lysate
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 50 seconds.
All lanes:
Immunoprecipitation - Anti-Ras antibody [EPR23474-20] (<a href='/en-us/products/primary-antibodies/ras-antibody-epr23474-20-ab275875'>ab275875</a>)
Predicted band size: 21 kDa
Observed band size: 21 kDa
false
- mIHC
Lab
Multiplex immunohistochemistry - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse stomach tissue staining VAChT with ab317452 at 1/1000 dilution, ab322711 anti-ALDH3A1 used at 1/500 dilution and ab275875 anti-Ras used at 1/2000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody.
Panel A : merged staining of anti-VAChT (green; Opal™520), anti-ALDH3A1 (gray; Opal™570) and anti-Ras (magenta; Opal™690) on mouse stomach.
Panel B : anti-VAChT staining nerve plexus in mouse stomach.
Panel C : ant-ALDH3A1 staining epithelium in mouse stomach.
Panel D : ant-Ras showed mainly membranous staining in mouse stomach.
Nuclear DNA was labeled with DAPI (shown in blue).
The section was incubated in three rounds of staining : in the order of ab317452, ab322711 and ab275875 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND®RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Ras with ab275875 at 1/2000 (0.338 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat colon. The section was incubated with ab275875 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
- IHC-P
Unknown
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling Ras with ab275875 at 1/2000 (0.338 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Mouse stomach. The section was incubated with ab275875 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
- Flow Cyt (Intra)
Unknown
Flow Cytometry (Intracellular) - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Ras with ab275875 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
A Goat anti rabbit IgG (Alexa Fluor®488, ab150077) at 1/2000 dilution was used as the secondary antibody.
- WB
Lab
Western blot - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration : 5% NFDM/TBST.
Exposure time : 15 seconds.
All lanes:
Western blot - Anti-Ras antibody [EPR23474-20] (<a href='/en-us/products/primary-antibodies/ras-antibody-epr23474-20-ab275875'>ab275875</a>) at 1/1000 dilution
Lane 1:
His-tagged human KRAS recombinant protein, 20 ng
Lane 2:
His-tagged human HRAS recombinant protein, 20 ng
Lane 3:
His-tagged human NRAS recombinant protein, 20 ng
Secondary
All lanes:
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Predicted band size: 21 kDa
false
- WB
Lab
Western blot - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration : 5% NFDM/TBST.
Exposure time : Lanes 1-2 : 26 seconds; Lane3 : 10 seconds.
All lanes:
Western blot - Anti-Ras antibody [EPR23474-20] (<a href='/en-us/products/primary-antibodies/ras-antibody-epr23474-20-ab275875'>ab275875</a>) at 1/1000 dilution
Lane 1:
Human colon tissue lysate at 40 µg
Lane 2:
Human liver tissue lysate at 40 µg
Lane 3:
Human stomach tissue lysate at 40 µg
Secondary
All lanes:
VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDa
false
- WB
Lab
Western blot - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration : 5% NFDM/TBST
Exposure time : 26 seconds.
All lanes:
Western blot - Anti-Ras antibody [EPR23474-20] (<a href='/en-us/products/primary-antibodies/ras-antibody-epr23474-20-ab275875'>ab275875</a>) at 1/1000 dilution
Lane 1:
HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 40 µg
Lane 2:
HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 40 µg
Lane 3:
A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 40 µg
Lane 4:
Human brain tissue lysate at 40 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDa
false
- WB
Lab
Western blot - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration : 5% NFDM/TBST
Exposure time : 3 seconds.
All lanes:
Western blot - Anti-Ras antibody [EPR23474-20] (<a href='/en-us/products/primary-antibodies/ras-antibody-epr23474-20-ab275875'>ab275875</a>) at 1/1000 dilution
Lane 1:
Mouse colon tissue lysate at 40 µg
Lane 2:
Mouse stomach tissue lysate at 40 µg
Lane 3:
Rat colon tissue lysate at 40 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDa
false
- WB
Lab
Western blot - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (AB275884)
This data was developed using ab275875, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration : 5% NFDM/TBST.
Exposure time : Lanes 1-3 : 26 seconds; Lane 4 : 70 seconds.
All lanes:
Western blot - Anti-Ras antibody [EPR23474-20] (<a href='/en-us/products/primary-antibodies/ras-antibody-epr23474-20-ab275875'>ab275875</a>) at 1/1000 dilution
Lane 1:
C6 (rat glial tumor glial cell) whole cell lysate at 20 µg
Lane 2:
RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 3:
PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Lane 4:
NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDa
false
Related conjugates and formulations (2)
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Anti-Ras antibody [EPR23474-20]
-
519 Alexa Fluor® 488
Alexa Fluor® 488 Anti-Ras antibody [EPR23474-20]
Reactivity data
Product details
ab275884 is the carrier-free version of ab275875.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
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 batch production
For more information, read more on recombinant antibodies.
Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties and storage information
Form
Purification technique
Storage buffer
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com