Antibodies, Proteins, Kits and Reagents for Life Science

311 587 ₸ Product size

100 µl 311 587 ₸ 10 µl 127 315 ₸

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Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP197Y] to Erk1 (pT202/pY204) + Erk2 (pT185/pY187)
Suitable for: WB, IP, Dot blot, ELISA
Reacts with: Mouse, Rat, Human

Product name

Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y]
See all Erk1 (pT202/pY204) + Erk2 (pT185/pY187) primary antibodies
Description

Rabbit monoclonal [EP197Y] to Erk1 (pT202/pY204) + Erk2 (pT185/pY187)
Host species

Rabbit
Specificity

This antibody detects Erk1 phosphorylated at Threonine 202 and Tyrosine 204 and Erk2 phosphorylated at Threonine 185 and Tyrosine 187.

Tested Applications & Species

Application	Species
IP	Rat Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: A431 (treated with EGF), SH-SY5Y (treated with NGF), PC-12 (treated with NGF), HeLa and NIH/3T3 whole cell lysates. IP: A431 (treated with EGF) and PC-12 (treated with NGF) whole cell lysates.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP197Y
Isotype

IgG
Research areas

Dot Blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

Dot blot analysis of single phospho peptide pT202 (Lane 1), single phospho peptide pY204 (Lane 2), double phospho peptide pT202/pY204 (Lane 3) and non-phospho peptide (Lane 4) labelling Erk1 (pT202/pY204) + Erk2 (p185/pY187) with ab76299 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/1000.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/5000 dilution

Lane 1 : A431 whole cell lysate - untreated
Lane 2 : A431 whole cell lysate - treated with 100ng/ml EGF for 10 minutes

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 43, 41 kDa
Observed band size: 44 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution

Lane 1 : SH-SY5Y whole cell lysate - untreated
Lane 2 : SH-SY5Y whole cell lysate - treated with 100ng/ml NGF for 10 minutes

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 43, 41 kDa
Observed band size: 42,44 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution

Lane 1 : PC-12 whole cell lysate - untreated
Lane 2 : PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 minutes

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 43, 41 kDa
Observed band size: 42,44 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution

Lane 1 : HeLa whole cell lysate - untreated
Lane 2 : HeLa whole cell lysate - treated with Lambda Phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 43, 41 kDa
Observed band size: 42,44 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution + NIH/3T3 whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 43, 41 kDa
Observed band size: 42,44 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking and dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

ab76299 at 1/80 immunoprecipitating Erk1 (pT202/pY204) + Erk2 (pT185/pY187) in A431 whole cell lysate.

Lane 1 (input): A431 whole cell lysate - treated with 100ng/ml EGF for 10 min (10µg).

Lane 2 (+): ab76299 + A431 whole cell lysate - treated with 100ng/ml EGF for 10 min.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76299 in A431 whole cell lysate - treated with 100ng/ml EGF for 10 min.

For western blotting, ab76299 was used at a dilution of 1/1000 and a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG, was used as the secondary antibody (1/1500).

Blocking and dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

ab76299 at 1/80 immunoprecipitating Erk1 (pT202/pY204) + Erk2 (pT185/pY187) in PC-12 whole cell lysate.

Lane 1 (input): PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 min (10µg).

Lane 2 (+): ab76299 + PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 min.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76299 in PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 min.

For western blotting, ab76299 was used at a dilution of 1/1000 and a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG, was used as the secondary antibody (1/1500).

Blocking and dilution buffer: 5% NFDM/TBST.
ELISA - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

ELISA analysis of various phospho and non-phospho peptides (0-250 ng/ml) labelling Erk1 (pT202/pY204) + Erk2 (pT185/pY187) with ab76299 at a dilution of 1/1000. An Alkaline Phosphatase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).

ab76299 has stronger affinity for the double phospho peptide Y204/Y187 than to single phospho peptides T202 or Y204.
Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

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