Anti-Ras antibody [EPR23474-20] (ab275875)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23474-20] to Ras
- Suitable for: WB, IHC-P, IP, Flow Cyt
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Ras antibody [EPR23474-20]
See all Ras primary antibodies -
Description
Rabbit monoclonal [EPR23474-20] to Ras -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt MouseHumanIHC-P MouseRatHumanIP HumanWB MouseRatHuman -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HEK-293T, A431, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human brain, colon, liver and stomach tissue lysates; Mouse colon and stomach tissue lysates; rat colon tissue 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
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23474-20 -
Isotype
IgG -
Research areas
Images
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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.
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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.
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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.
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All lanes : Anti-Ras antibody [EPR23474-20] (ab275875) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : A431 (human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 4 : Human brain tissue lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 26 seconds.
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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.
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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.
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All lanes : Anti-Ras antibody [EPR23474-20] (ab275875) at 1/1000 dilution
Lane 1 : Human colon tissue lysate
Lane 2 : Human liver tissue lysate
Lane 3 : Human stomach tissue lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: Lanes 1-2: 26 seconds; Lane3: 10 seconds.
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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.
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All lanes : Anti-Ras antibody [EPR23474-20] (ab275875) at 1/1000 dilution
Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 2 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma ) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: Lanes 1-3: 26 seconds; Lane 4: 70 seconds.
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Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Ras with abab275875 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.
-
All lanes : Anti-Ras antibody [EPR23474-20] (ab275875) at 1/1000 dilution
Lane 1 : Mouse colon tissue lysate
Lane 2 : Mouse stomach tissue lysate
Lane 3 : Rat colon tissue lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 seconds.
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All lanes : Anti-Ras antibody [EPR23474-20] (ab275875) 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 (ab97051) at 1/100000 dilution
Predicted band size: 21 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds.
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