Antibodies, Proteins, Kits and Reagents for Life Science

251 280 ₸ Product size

100 µl 251 280 ₸

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Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E268] to ATF2 (phospho T71)
Suitable for: WB, IP, ICC/IF, Dot blot
Reacts with: Mouse, Human

Product name

Anti-ATF2 (phospho T71) antibody [E268]
See all ATF2 primary antibodies
Description

Rabbit monoclonal [E268] to ATF2 (phospho T71)
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IP	Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide corresponding to Human ATF2 aa 50-150.
Database link: P15336
Positive control
- IP: HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate. ICC/IF: HeLa cells treated with 250ng/ml anisomycin for 30min.
General notes
SAPK and p38 MAPK activate, in response to cellular stress, ATF2 by phosphorylating the protein at Thr69 and Thr71. Mutations of these sites result in the loss of stress induced transcription by ATF2.

Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Primary antibody notes

SAPK and p38 MAPK activate, in response to cellular stress, ATF2 by phosphorylating the protein at Thr69 and Thr71. Mutations of these sites result in the loss of stress induced transcription by ATF2.
Clonality

Monoclonal
Clone number

E268
Isotype

IgG
Research areas

Western blot - Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

All lanes : Anti-ATF2 (phospho T71) antibody [E268] (ab32019) at 0.526 µg/ml

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) grown in serum free media for overnight, whole cell lysate
Lane 2 : HeLa grown in serum free media for overnight, then treated with 250ng/ml Anisomycin for 30min, whole cell lysate
Lane 3 : HeLa grown in serum free media for overnight, then treated with 250ng/ml Anisomycin for 30min whole cell lysate. Then the membrane was incubated with alkaline phosphatase.

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/ml

Predicted band size: 54 kDa
Observed band size: 70 kDa
why is the actual band size different from the predicted?

Blocking/Diluting Buffer and concentration: 5% NFDM /TBST
Immunocytochemistry/ Immunofluorescence - Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

Immunocytochemistry of HeLa (Human epithelial cell line from cervix adenocarcinoma), prepared in FBS free medium overnight labeling ATF2 at 0.9 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. AlexaFluor^®488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at 1/500 . Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used as the counter stain at 2.5 μg/ml. DAPI was used for nuclear counter stain. Confocal image showing the expression was increased on HeLa cells, prepared in FBS free medium overnight, then treated with 250ng/ml anisomycin for 30min.
Western blot - Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

All lanes : Anti-ATF2 (phospho T71) antibody [E268] (ab32019) at 0.1 µg/ml

Lane 1 : Untreated NIH/3T3 (mouse embryo), prepared in FBS free medium overnight, whole cell lysate
Lane 2 : NIH/3T3, prepared in FBS free medium overnight, then treated with 10 µg/ml anisomycin for 30 minutes whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 54 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking and diluting buffer used was 2% BSA/TBST.
Immunoprecipitation - Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

Lane 1 (input): HeLa (human cervix adenocarcinoma) treated with 250ng/ml anisomycin for 30min whole cell lysate 10μg.
Lane 2 (+): HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32019 in HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate.

Ab32019 Immunoprecipitating ATF2 in Human Hela whole cell lysate. For western blotting ab32019 (1:1000) was used to confirm successful immunoprecipitation. Blocking and diluting buffer used was 5% NFDM/TBST.

All lanes : Anti-ATF2 (phospho T71) antibody [E268] (ab32019) at 0.47 µg/ml

Lane 1 : HeLa (human cervix adenocarcinoma) treated with 250ng/ml anisomycin for 30min whole cell lysate at 10 µg
Lane 2 : HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab32019 in HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 dilution

Exposure time: 3 minutes
Dot Blot - Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

Dot blot analysis of ATF2 (phospho T71) peptide (Lane 1) and ATF2 non-phospho peptide (Lane 2) using ab32019 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

Exposure time: 3 minutes

Blocking and Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

All lanes : Anti-ATF2 (phospho T71) antibody [E268] (ab32019) at 0.1 µg/ml

Lane 1 : HeLa (human cervix adenocarcinoma), prepared in FBS free medium overnight, whole cell lysate
Lane 2 : HeLa, prepared in FBS free medium overnight, then treated with 250ng/ml anisomycin for 30 minutes whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 54 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking and diluting buffer used was 2% BSA/TBST .
The molecular weight is 70kD, consistent with the literature (PMID: 24223142).
Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

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