Overview

  • Product name

  • Description

    Rabbit polyclonal to ATF2
  • Host species

    Rabbit
  • Specificity

    ATF2 antibody detects endogenous levels of ATF2 protein around Threonine 71 or 53.
  • Tested applications

    Suitable for: WB, IHC-P, ELISA, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    The antiserum was produced against synthesized non-phosphopeptide derived from human ATF2 around the phosphorylation site of Threonine 71 or 53.

Properties

Applications

Our Abpromise guarantee covers the use of ab47476 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/1000. Predicted molecular weight: 52 kDa.
IHC-P Use at an assay dependent concentration.
ELISA 1/20000.
IP Use at an assay dependent concentration.

Target

  • Function

    Transcriptional activator, probably constitutive, which binds to the cAMP-responsive element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. Interaction with JUN redirects JUN to bind to CRES preferentially over the 12-O-tetradecanoylphorbol-13-acetate response elements (TRES) as part of an ATF2/JUN complex.
  • Tissue specificity

    Abundant expression seen in the brain.
  • Sequence similarities

    Belongs to the bZIP family. ATF subfamily.
    Contains 1 bZIP domain.
    Contains 1 C2H2-type zinc finger.
  • Post-translational
    modifications

    Phosphorylation of Thr-69 and Thr-71 by MAPK14 causes increased transcriptional activity. Also phosphorylated and activated by JNK.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Activating transcription factor 2 antibody
    • Activating transcription factor 2 splice variant ATF2 var2 antibody
    • ATF 2 antibody
    • Atf-2 antibody
    • Atf2 antibody
    • ATF2 protein antibody
    • ATF2_HUMAN antibody
    • cAMP Response Element Binding Protein 2 antibody
    • cAMP response element binding protein CRE BP1 antibody
    • cAMP response element-binding protein CRE-BP1 antibody
    • cAMP responsive element binding protein 2, formerly antibody
    • cAMP-dependent transcription factor ATF-2 antibody
    • cAMP-responsive element-binding protein 2 antibody
    • CRE BP1 antibody
    • CRE-BP antibody
    • CREB 2 antibody
    • CREB-2 antibody
    • CREB2 antibody
    • CREBP1 antibody
    • Cyclic AMP dependent transcription factor ATF 2 antibody
    • Cyclic AMP-dependent transcription factor ATF-2 antibody
    • Cyclic AMP-responsive element-binding protein 2 antibody
    • D130078H02Rik antibody
    • D18875 antibody
    • HB 16 antibody
    • HB16 antibody
    • Histone acetyltransferase ATF2 antibody
    • MGC105211 antibody
    • MGC105222 antibody
    • MGC111558 antibody
    • MGC142504 antibody
    • mXBP antibody
    • MXBP protein antibody
    • Tg(Gzma-Klra1)7Wum antibody
    • TREB 7 antibody
    • TREB7 antibody
    see all

Images

  • All lanes : Anti-ATF2 antibody (ab47476) at 1/500 dilution

    Lane 1 : extracts from HeLa cells.
    Lane 2 : extracts from HeLa cells preincubated with immunizing peptide

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

  • Immunohistochemical analysis of paraffin-embedded breast carcinoma. Left: Using ATF2 antibody; Right: The same antibody preincubated with synthesized peptide.

References

This product has been referenced in:

  • Zhou Y  et al. Ligustilide attenuates nitric oxide-induced apoptosis in rat chondrocytes and cartilage degradation via inhibiting JNK and p38 MAPK pathways. J Cell Mol Med 23:3357-3368 (2019). Read more (PubMed: 30770640) »
  • Liu F  et al. The role of NF-?B-mediated JNK pathway in cognitive impairment in a rat model of sleep apnea. J Thorac Dis 10:6921-6931 (2018). Read more (PubMed: 30746238) »
See all 8 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question

Hello , Here is the complain of two antibodies .

1) Abcam catalogue no:

Ab31419, Ab47476

2) a) Abcam order number (not required):

1148646


b) Abcam product lot number:

c-Jun> GR50836-9, ATF-2> GR72060-1

3) Description of the problem (high background, wrong band size, more bands, no band etc):

No bands

4) Antibody storage conditions (temperature/reconstitution etc):

-20°C


5) a)Sample (Species):


c-Jun and ATF2 from HeLa


b) Type of sample (Cell extract/Nuclear extract/Purified protein/Recombinant protein etc).:


Whole cell extract

6) a) Sample preparation:

Lysis buffer



RIPA





Protease inhibitors



AEBSF, 2 mM
Aprotinin, 0.3 μM
Bestatin, 130 μM
EDTA, 1 mM
E-64, 14 μM
Leupeptin, 1 μM

PMSF, 20 mM





Phosphatase inhibitors



-





Reducing agent



SDS and β-ME





Boiling for ≥5 min?



5 minutes







b) Amount of protein loaded:







Protein loaded ug/lane or cells/lane



5 μg/lane







7) a) Electrophoresis/Gel conditions:







Reducing or Non Reducing gel



Reducing





Percentage of gel



15%





Volts applied



75





Time applied



3 hours

















b) Transfer or blocking conditions:







Type of membrane



PVDF





Protein transfer verified



Ponceau, β-actin





Blocking agent and concentration



3% BSA





Blocking time



30 minutes





Blocking temperature



25°C

















8) Primary antibody (If more than one was used, describe in “additional notes”):









Concentration or dilution



1/1000





Diluent buffer



1% BSA + 0.05% Tween-20 in 1X PBS





Incubation time



2 hours





Incubation temperature



25°C





Washing: Buffer Used



0.05% Tween-20 in 1X PBS





Number of washes



3







9) Secondary antibody:









Species



Goat





Reacts against



Rabbit





Concentration or dilution



1/10,000





Diluent buffer



1% BSA + 0.05% Tween-20 in 1X PBS





Incubation time



2 hours





Incubation temperature:



25°C





Fluorochrome or enzyme conjugate



HRP





Washing: Buffer Used



0.05% Tween-20 in 1X PBS





Number of washes



3







10) Detection method (ECL, ECL plus, etc.):









ECL





11) Positive and negative controls used:









Positive control



β-actin





Negative control











12) a) How many times you have run this staining?







Twice







b) Do you obtain the same results every time?







No







C) What steps have you altered to try and optimize the use of this antibody?







Reducing blocking, primary and secondary incubation times while attempting the second time. Washing was made slightly rigorous as compared to he first time. Performed a Dot blot to check for performance of secondary antibody and it works fine. Increased amount of loaded protein to 10 μg/lane.







Regards

Read More
Answer

Thank you for your enquiry regarding ab31419 and ab47476; and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with these two antibodies.
I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.
After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:
1) The loaded amount of protein seems to be a bit low: 5 μg/lane, normally we would suggest loading at least 25-30 μg/lane protein per lane.
2) Since c-Jun and ATF2 are both mainly present in the nuclei, it would be worth preparing nuclear fraction (rather than whole cell lysate) to enrich the target proteins in the sample if it is possible and apply them for WB.
3) Customer may also need to test different dilutions (1/500 or even 1/250) to make sure that the experiment is carried out under the optimal conditions. This is even more important since only 5 μg instead of 30 μg lysate was loaded.
I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (endothelial)
Loading amount
5000 cells
Specification
endothelial
Treatment
different shear stress exposure for 24 hours
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

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Verified customer

Submitted Mar 01 2010

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