Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-Ras antibody [EPR23474-20] - BSA and Azide free
See all Ras primary antibodies
Description

Rabbit monoclonal [EPR23474-20] to Ras - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
IHC-P	Rat
IP	Human
WB	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
ab275884 is the carrier-free version of ab275875. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab275884 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C.
Storage buffer

Constituent: 100% PBS
Carrier free

Yes
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] - 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.
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.
Flow Cytometry - 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.

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] - BSA and Azide free (ab275884)

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 : 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

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.
Western blot - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (ab275884)

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

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.
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.
Western blot - Anti-Ras antibody [EPR23474-20] - BSA and Azide free (ab275884)

All lanes : Anti-Ras antibody [EPR23474-20] (ab275875) at 1/1000 dilution

Lanes 1 & 3 : 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 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

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.
Flow Cytometry - 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.

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] - BSA and Azide free (ab275884)

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

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.
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 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] - BSA and Azide free (ab275884)

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

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.
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.
Anti-Ras antibody [EPR23474-20] - BSA and Azide free (ab275884)