Overview

  • Product name

    Anti-Transcription factor AP-2-alpha antibody
    See all Transcription factor AP-2-alpha primary antibodies
  • Description

    Rabbit polyclonal to Transcription factor AP-2-alpha
  • Host species

    Rabbit
  • Specificity

    ab52222 detects endogenous levels of total Transcription factor AP-2-alpha protein.
  • Tested applications

    Suitable for: ChIP, ICC/IF, EMSA, WB, ELISA, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Chicken
  • Immunogen

    Synthetic peptide corresponding to Human Transcription factor AP-2-alpha.
    Database link: P05549

  • Positive control

    • WB: Extracts from COLO 205 cells. IHC-P: Mouse embryonic tissue. Human breast carcinoma tissue. ICC/IF: SiHa cells.

Properties

  • 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: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride

    Without Mg+2 and Ca+2
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab52222 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
ChIP Use at an assay dependent concentration.
ICC/IF 1/1000. (see Abreview)
EMSA Use at an assay dependent concentration.
WB 1/500 - 1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
ELISA 1/10000.
IHC-P Use a concentration of 5 µg/ml.
EMSA Use at an assay dependent concentration. PubMed: 20671194

Target

  • Function

    Sequence-specific DNA-binding protein that interacts with inducible viral and cellular enhancer elements to regulate transcription of selected genes. AP-2 factors bind to the consensus sequence 5'-GCCNNNGGC-3' and activate genes involved in a large spectrum of important biological functions including proper eye, face, body wall, limb and neural tube development. They also suppress a number of genes including MCAM/MUC18, C/EBP alpha and MYC. AP-2-alpha is the only AP-2 protein required for early morphogenesis of the lens vesicle.
  • Involvement in disease

    Defects in TFAP2A are the cause of branchiooculofacial syndrome (BOFS) [MIM:113620]; also known as branchial clefts with characteristic facies, growth retardation, imperforate nasolacrimal duct, and premature aging or lip pseudocleft-hemangiomatous branchial cyst syndrome. BOFS is a rare autosomal dominant cleft palate craniofacial disorder with variable expressivity. The major features include cutaneous anomalies, ocular anomalies, characteristic facial appearance (malformed pinnae, oral clefts), and, less commonly, renal and ectodermal (dental and hair) anomalies.
  • Sequence similarities

    Belongs to the AP-2 family.
  • Domain

    The WW-binding motif mediates interaction with WWOX.
  • Post-translational
    modifications

    Sumoylated on Lys-10; which inhibits transcriptional activity.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Activating enhancer binding protein 2 alpha antibody
    • Activating enhancer-binding protein 2-alpha antibody
    • Activator protein 2 antibody
    • AP 2 transcription factor antibody
    • AP 2alpha antibody
    • AP-2 antibody
    • AP-2 transcription factor antibody
    • AP2 antibody
    • AP2 Transcription Factor antibody
    • AP2-alpha antibody
    • AP2A_HUMAN antibody
    • AP2TF antibody
    • BOFS antibody
    • FLJ51761 antibody
    • TFAP 2 antibody
    • TFAP 2A antibody
    • TFAP2 antibody
    • TFAP2A antibody
    • Transcription factor AP 2 alpha (activating enhancer binding protein 2 alpha) antibody
    • Transcription factor AP-2-alpha antibody
    • Transcription factor AP2 alpha antibody
    see all

Images

  • SiHa (Human cervical cell lines) cells stained for Transcription factor AP-2-alpha (Green) using ab52222 (1/500 dilution) in ICC/IF. Secondary used is an Alexa-Fluor®488-conjugated Goat Anti-Rabbit IgG (H+L).

    See Abreview

  • Immunohistochemical analysis of formaldehyde-fixed paraffin-embedded murine embryonic tissue sections, labelling Transcription factor AP-2-alpha with ab52222 at a dilution of 1/3000 incubated for 2 hours at 21°C in TBS, BSA and azide diluent. Heat mediated antigen retrival was with citric acid. Blocking was with 1% BSA incubated for 10 minutes at 21°C. Secondary was a goat anti-rabbit polyclonal biotin conjugate at 1/300. In this day 18 embryo, nuclear positivity was also detected in spinal cord, ganglia, kidney tubules and the keratinised portion of the stomach.

    See Abreview

  • All lanes : Anti-Transcription factor AP-2-alpha antibody (ab52222) at 1/500 dilution

    Lane 1 : COLO 205 (Human colon adenocarcinoma cell line) extracts
    Lane 2 : COLO 205 (Human colon adenocarcinoma cell line) extracts with immunizing peptide

    Predicted band size: 48 kDa
    Observed band size: 48 kDa

  • ab52222 was used in an Electrophoretic Mobility Shift Assay (EMSA) to supershift the protein-DNA complex.

    Radiolabeled, double-stranded DNA oligonucleotides (10.000 cpm per lane) harbouring a binding site for Transcription factor AP-2-alpha were incubated with each 2 µg of nuclear extract (NE) from HeLa and Caski cells, respectively. Samples were incubated for 30 minutes at room temperature to allow the formation of protein-DNA complexes. 2 µg of anti-Transcription factor AP-2-alpha antibody were added to the samples (as indicated) and incubated for further 60 minutes at 4°C. Samples were separated in a 5.5% PAGE for 30 minutes at 280 V and further 75 minutes at 350 V. The gel was dried under vacuum and for autoradiography a X-ray film was exposed with an intensifying screen for 2 days at -80°C. Specific protein-DNA complexes were quantitatively supershifted with antibody anti-AP-2 alpha (ab52222), verifying the binding of Transcription factor AP-2-alpha to the DNA oligonucleotide.

  • IHC image of Transcription factor AP-2-alpha staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab52222, 5 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

References

This product has been referenced in:

  • Xu J  et al. miR-876-5p suppresses breast cancer progression through targeting TFAP2A. Exp Ther Med 18:1458-1464 (2019). Read more (PubMed: 31316633) »
  • Chen XF  et al. An Osteoporosis Risk SNP at 1p36.12 Acts as an Allele-Specific Enhancer to Modulate LINC00339 Expression via Long-Range Loop Formation. Am J Hum Genet 102:776-793 (2018). Read more (PubMed: 29706346) »
See all 15 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Embryo skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Embryo skin
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Sep 21 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Embryonic spinal cord)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Embryonic spinal cord
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Sep 21 2015

Answer

Thank you for contacting us.

The exact immunogen sequence is not available to us as the originating laboratory considers it proprietary, but we at least know that it is within the final 30 amino acids of the C terminus of the human protein, Uniprot accession P05549 (http://www.uniprot.org/uniprot/P05549).

AMDKMYLSNNPNSHTDNNAKSSDKEEKHRK.

This may be sufficient for your alignments. If not, please send the sequences or accession numbers of the proteins you need to align, and I will forward them to the originating lab for alignment to obtain the percentage identities.

Read More

Answer

Thank you for your inquiry.

I am happy to confirm that ab500 (ChIP Kit) is suitable for this antibody since it can be used with rabbit antibodies.

https://www.abcam.com/index.html?datasheet=500 (or use the following: https://www.abcam.com/index.html?datasheet=500).

This kit has not been experimentally tested with ab52222, but since ab52222 is tested and guaranteed for CHIP, the combination of this kit and ab52222 is also guaranteed.

I hope this information is helpful and wish you good luck for your experiments.

Read More

Answer

Thank you for contacting us.

Reactivity with chicken AP2-2 alpha is predicted based on 100% conservation of the immunogen in the chicken protein NP_990425.1.

However, we do not have data confirming this, and reactivity with chicken AP2-2 alpha is not guaranteed.

Please do not hesitate to contact us if you need any more advice or information.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human cervical cell lines)
Permeabilization
Yes - triton
Specification
human cervical cell lines
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 14 2011

Application
ChIP
Sample
Human Cell lysate - nuclear (Cervical cancer cell lines)
Negative control
Human primary keratinocytes
Specification
Cervical cancer cell lines
Detection step
Semiquantitative PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Positive control
Tumor cell lines Hela

Abcam user community

Verified customer

Submitted Jan 03 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human keratinocytes)
Permeabilization
Yes - 0.1% triton
Specification
human keratinocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 20 2010

Application
Western blot
Sample
Human Cell lysate - whole cell (Human keratinocytes)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
50 µg
Specification
Human keratinocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 16 2010

Application
Western blot
Sample
Human Cell lysate - whole cell (Hybrid fibroblast-cervical tumor keratinocyte)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
100 µg
Specification
Hybrid fibroblast-cervical tumor keratinocyte
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 10 2010

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