Antibodies, Proteins, Kits and Reagents for Life Science

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

Application	Species
Flow Cyt	Mouse Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23474-20
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR23474-20] (ab275875)

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.
Immunoprecipitation - Anti-Ras antibody [EPR23474-20] (ab275875)

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.
Flow Cytometry - Anti-Ras antibody [EPR23474-20] (ab275875)

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.
Western blot - Anti-Ras antibody [EPR23474-20] (ab275875)

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 kDa

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 26 seconds.
Immunoprecipitation - Anti-Ras antibody [EPR23474-20] (ab275875)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR23474-20] (ab275875)

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.
Western blot - Anti-Ras antibody [EPR23474-20] (ab275875)

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 kDa

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Exposure time: Lanes 1-2: 26 seconds; Lane3: 10 seconds.
Flow Cytometry - Anti-Ras antibody [EPR23474-20] (ab275875)

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.
Western blot - Anti-Ras antibody [EPR23474-20] (ab275875)

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 kDa

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Exposure time: Lanes 1-3: 26 seconds; Lane 4: 70 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR23474-20] (ab275875)

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.
Western blot - Anti-Ras antibody [EPR23474-20] (ab275875)

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 kDa

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 3 seconds.
Western blot - Anti-Ras antibody [EPR23474-20] (ab275875)

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 kDa

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Exposure time: 15 seconds.
Anti-Ras antibody [EPR23474-20] (ab275875)