Overview

  • Product name
    Anti-PAX9 antibody [7C2]
    See all PAX9 primary antibodies
  • Description
    Rat monoclonal [7C2] to PAX9
  • Host species
    Rat
  • Specificity
    Does not cross react with other PAX proteins
  • Tested applications
    Suitable for: IP, IHC-Fr, ICC/IF, IHC-P, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant PAX9-MBP fusion protein (Mouse)

Properties

Applications

Our Abpromise guarantee covers the use of ab28538 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
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 10 µg/ml.
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 38 kDa.
Flow Cyt Use 1µg for 106 cells.

ab18407 - Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    Transcription factor required for normal development of thymus, parathyroid glands, ultimobranchial bodies, teeth, skeletal elements of skull and larynx as well as distal limbs.
  • Involvement in disease
    Defects in PAX9 are the cause of tooth agenesis selective type 3 (STHAG3) [MIM:604625]. A form of selective tooth agenesis, a common anomaly characterized by the congenital absence of one or more teeth. Selective tooth agenesis without associated systemic disorders has sometimes been divided into 2 types: oligodontia, defined as agenesis of 6 or more permanent teeth, and hypodontia, defined as agenesis of less than 6 teeth. The number in both cases does not include absence of third molars (wisdom teeth).
  • Sequence similarities
    Contains 1 paired domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Paired box 9 antibody
    • Paired box gene 9 antibody
    • Paired box homeotic gene 9 antibody
    • Paired box protein 9 antibody
    • Paired box protein Pax 9 antibody
    • Paired box protein Pax-9 antibody
    • Paired box protein Pax9 antibody
    • Paired domain gene 9 antibody
    • PAX 9 antibody
    • PAX9 antibody
    • PAX9_HUMAN antibody
    • STHAG3 antibody
    see all

Images

  • ab28538 staining PAX9 in mouse E11.5 embryo tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with 2% paraformaldehyde in PBS for 2 hours at 4°C then placed in 30% sucrose overnight at 4°C prior to embedding in OCT using liquid nitrogen. Tissue was blocked with 1X PBS + 1.5% Milk + 1.5% BSA + 0.1% Triton X-100 for 12 hours at 4°C. Samples were incubated with primary antibody (1/500 in blocking buffer) for 1 hour at 23°C. An Alexa Fluor® 568-conjugated goat anti-rat IgG polyclonal (1/250) was used as the secondary antibody.

    See Abreview

  • Staining of PAX9 protein in human oesophageal epithelium. Magnification x200
  • All lanes : Anti-PAX9 antibody [7C2] (ab28538)

    Lane 1 : mouse oesophageal protein extract
    Lane 2 : human oesophageal protein extract

    Predicted band size: 38 kDa
    Observed band size: 38 kDa

  • ICC/IF image of ab28538 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28538, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HepG2 cells stained with ab28538 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab28538, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was goat anti-rat DyLight® 488  (IgG H+L) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG1, kappa monoclonal [RTK2071] (ab18412, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:
  • Feng J  et al. BMP signaling orchestrates a transcriptional network to control the fate of mesenchymal stem cells in mice. Development 144:2560-2569 (2017). Read more (PubMed: 28576771) »
  • Sherwood RI  et al. Transcriptional dynamics of endodermal organ formation. Dev Dyn 238:29-42 (2009). ICC/IF ; Mouse . Read more (PubMed: 19097184) »
See all 5 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (E11.5 embryos)
Permeabilization
No
Specification
E11.5 embryos
Blocking step
PBSMBT (1X PBS, 1.5% Milk, 1.5% BSA, 0.1% Triton X-100) as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 4°C
Fixative
Paraformaldehyde

Mr. William Wood

Verified customer

Submitted Apr 30 2014

Question
Answer

Here is the IHC protocol I received from the laboratory that validated the antibody for this application. Please let me know if you have any questions. As I mentioned in my previous e-mail, if your protocol is similar to this one, I will be happy to discuss a replacement.

Serial sections (5 mm) were

mounted onto poly-L-lysine-coated slides, baked at

65°C overnight, deparaffinized and dehydrated according

to standard protocols. For antigen retrieval, specimens

were boiled for 4 min in a pressure cooker in

citrate buffer (10 mM, pH 6.0). Non-specific binding of

the primary antibodies was reduced by pre-incubation

of the slides with 4% dried skimmed milk in antibody

diluent. Immunostaining was performed by the streptavidin–

alkaline phosphatase technique using the Dako

ChemMate Detection Kit (DAKO, Denmark) according

to the manufacturer’s instruction, and using Fast

Red as a colour substrate. Sections were counterstained

with haematoxylin. Only distinct nuclear

staining of PAX9 was considered positive and staining

intensity was not graded in this study. Immunoreactive

cells were estimated as a percentage of the total number

of tumour cells and dysplastic cells, respectively.

PAX9 expression in dysplasias and in invasive carcinomas

was classified as reduced when the fraction of

PAX9-positive cells was smaller than 50%, and

increased when the fraction of PAX9-positive cells

was greater than 95%.

Read More

Answer

Thank you for your enquiry and your interest.

We have several antibodies against PAX9 but none of them have been tested specifically in ChIP application. Here are some which recognize mouse (murine) PAX9 and detect the native form of the protein so there is a high chance that they work in ChIP as well: ab28538, ab84830, ab85168.

I could certainly offer a Testing Discount code for your customer if she would liek to test one of them. The Terms and Conditions of this offer can be found at: https://www.abcam.com/collaborationdiscount.

Please do let me know if your customer is interested in this offer. I look forward to hearing from you soon.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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