Overview

  • Product name
  • Description
    Goat polyclonal to PAX3
  • Host species
    Goat
  • Tested applications
    Suitable for: WB, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Dog
  • Immunogen

    Synthetic peptide: TTLAGAVPRMM, corresponding to N terminal amino acids 2-12 of PAX3.

  • Positive control
    • Recombinant Human PAX3 protein (ab114320) can be used as a positive control in WB. Human squamous epithelium. Human duodenum lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab15717 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 Use a concentration of 0.3 - 1 µg/ml. Predicted molecular weight: 53 kDa.Can be blocked with Human PAX3 peptide (ab23270).
IHC-P Use a concentration of 3 µg/ml.
IP Use a concentration of 2.5 µg/ml.

Target

  • Function
    Probable transcription factor associated with development of alveolar rhabdomyosarcoma.
  • Involvement in disease
    Defects in PAX3 are the cause of Waardenburg syndrome type 1 (WS1) [MIM:193500]. WS1 is an autosomal dominant disorder characterized by wide bridge of nose owing to lateral displacement of the inner canthus of each eye (dystopia canthorum), pigmentary disturbances such as frontal white blaze of hair, heterochromia of irides, white eyelashes, leukoderma and sensorineural deafness. The syndrome shows variable clinical expression and some affected individuals do not manifest hearing impairment.
    Defects in PAX3 are the cause of Waardenburg syndrome type 3 (WS3) [MIM:148820]; also known as Klein-Waardenburg syndrome or Waardenburg syndrome with upper limb anomalies or white forelock with malformations. WS3 is a very rare autosomal dominant disorder, which shares many of the characteristics of WS1. Patients additionally present with musculoskeletal abnormalities.
    Defects in PAX3 are the cause of craniofacial-deafness-hand syndrome (CDHS) [MIM:122880]. CDHS is thought to be an autosomal dominant disease which comprises absence or hypoplasia of the nasal bones, hypoplastic maxilla, small and short nose with thin nares, limited movement of the wrist, short palpebral fissures, ulnar deviation of the fingers, hypertelorism and profound sensory-neural deafness.
    Defects in PAX3 are a cause of rhabdomyosarcoma type 2 (RMS2) [MIM:268220]. It is a form of rhabdomyosarcoma, a highly malignant tumor of striated muscle derived from primitive mesenchimal cells and exhibiting differentiation along rhabdomyoblastic lines. Rhabdomyosarcoma is one of the most frequently occurring soft tissue sarcomas and the most common in children. It occurs in four forms: alveolar, pleomorphic, embryonal and botryoidal rhabdomyosarcomas. Note=A chromosomal aberration involving PAX3 is found in rhabdomyosarcoma. Translocation (2;13)(q35;q14) with FOXO1. The resulting protein is a transcriptional activator.
    Note=A chromosomal aberration involving PAX3 is a cause of rhabdomyosarcoma. Translocation t(2;2)(q35;p23) with NCOA1 generates the NCOA1-PAX3 oncogene consisting of the N-terminus part of PAX3 and the C-terminus part of NCOA1. The fusion protein acts as a transcriptional activator. Rhabdomyosarcoma is the most common soft tissue carcinoma in childhood, representing 5-8% of all malignancies in children.
  • Sequence similarities
    Belongs to the paired homeobox family.
    Contains 1 homeobox DNA-binding domain.
    Contains 1 paired domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • CDHS antibody
    • HUP 2 antibody
    • HUP2 antibody
    • MGC120381 antibody
    • MGC120382 antibody
    • MGC120383 antibody
    • MGC120384 antibody
    • MGC134778 antibody
    • Paired box 3 antibody
    • Paired box gene 3 antibody
    • Paired box homeotic gene 3 antibody
    • Paired box protein Pax 3 antibody
    • Paired box protein Pax-3 antibody
    • Paired box protein Pax3 antibody
    • Paired domain gene 3 antibody
    • Paired domain gene HuP2 antibody
    • PAX 3 antibody
    • Pax3 antibody
    • PAX3/FKHR fusion gene antibody
    • PAX3_HUMAN antibody
    • Sp antibody
    • splotch antibody
    • Waardenburg syndrome 1 antibody
    • WS 1 antibody
    • WS1 antibody
    • WS3 antibody
    see all

Images

  • ab15717 staining PAX3 in murine striated muscle tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using citrated buffer pH 6. Samples were then blocked with 1% BSA for 30 minutes at 25°C and then incubated with ab15717 at a 1/100 dilution for 18 hours at 25°C. The secondary used was an undiluted HRP conjugated goat polyclonal.

    See Abreview

  • ab15717 (3µg/ml) staining of paraffin embedded Human Oesophagus. The tissue sections subjected to antigen retrieval by microwave in Tris/EDTA buffer pH 9.0. The HRP-staining procedure was used for detection.

  • Anti-PAX3 antibody (ab15717) at 0.3 µg/ml + Human duodenum lysate in RIPA buffer at 35 µg

    Developed using the ECL technique.

    Predicted band size: 53 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?

References

This product has been referenced in:
  • Sun L  et al. Mir193b-365 is essential for brown fat differentiation. Nat Cell Biol 13:958-65 (2011). WB . Read more (PubMed: 21743466) »
  • Dissanayake SK  et al. Wnt5A regulates expression of tumor-associated antigens in melanoma via changes in signal transducers and activators of transcription 3 phosphorylation. Cancer Res 68:10205-14 (2008). WB ; Human . Read more (PubMed: 19074888) »
See all 2 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Chicken Cell (Primary culture of chicken muscle cells)
Permeabilization
Yes - Triton X-100
Specification
Primary culture of chicken muscle cells
Blocking step
BSA + Milk + Serum as blocking agent for 20 minute(s) · Concentration: 2.0% · Temperature: 27°C
Fixative
Formaldehyde

Dr. Marcelo Pelajo Machado

Verified customer

Submitted Jul 21 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Chicken Tissue sections (Head of chicken embryo)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9.0
Permeabilization
No
Specification
Head of chicken embryo
Blocking step
BSA + Milk + Serum as blocking agent for 20 minute(s) · Concentration: 2.0% · Temperature: 27°C
Fixative
Formaldehyde

Dr. Marcelo Pelajo Machado

Verified customer

Submitted Jul 21 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (brain)
Permeabilization
No
Specification
brain
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 04 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Striated muscle)
Specification
Striated muscle
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrated Buffer, pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 22 2012

Answer

Thank you for your enquiry and your interest in our products. This product (ab15717) is expected to recognize all 7 isoforms based on the immunizing peptide. However, in WB we only see a 30-35 kDa band in human kidney lysates. I can confirm that ab15717 works in IHC-P as well (tested on formalin-fixed paraffin embedded human oesophagus tissue). In order to be able to distinguish between the different isoforms, Iso-Electric Focusing (IEF) technique can be applied. If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Question
Answer

Thank you for contacting us. The protocol used to test this antibody in IHC-P is as follows: *Buffers* Tris/ EDTA buffer at pH 9.0:
-Tris 1.21 g
-EDTA 0.37 g
-Distilled water 1000 ml (100 ml to make 10x, 50 ml to make 20x)
Mix to dissolve. pH is usually at 9.0. Add 0.5 ml of Tween 20 and mix well.
Store at room temperature for 3 months or at 4°C for longer storage. *1.1 Deparaffinise and Rehydrate Tissue Sections. * - Remove wax and bring the sections to water. *1.2 **Microwave Antigen Retrieval.* - Place slides in a microwaveable pot containing Tris/ EDTA buffer at pH 9.0, ensure slides are completely covered with solution. - Put pot and slides in microwaveable pressure cooker and place in microwave. - When the cooker has been heated up to pressure, “cook” for 1 - 3 minutes (as appropriate). - Remove the pressure cooker and allow to cool. - When the pressure has dropped, remove the pot containing the slides, and allow the slides to cool/rest. - Wash for a minimum of 5 minutes in tap water. - Wash for 5 minutes in distilled water. *1.3 **Blocking Endogenous Peroxidase.* - Treat sections with a H202 solution ( e.g. 1ml H202 20% in 250ml dH20) and incubate for 15. - Wash slides in PBS *1.4 **Primary Antibody Incubation.* - Apply primary antibody diluted to a concentration of 3ug/mL using 10% NHS in PBS. - Incubate slides at room temperature in a humidity staining chamber for 30 minutes. - Wash in PBS for 2 x 5 minutes. *1.5 **Secondary Antibody Incubation.* - Apply secondary antibody (anti-goat HRP) and incubate slides at room temperature in a humidity staining chamber for 30 minutes. - Wash in PBS for 2 x 5 minutes. *1.6 **Detection.* - Add DAB solution for 5 minutes. - Replace PBS with distilled water to wash for at least 15 minutes (change water frequently). - Counterstain with haematoxylin. Wash in tap water for at least 15 minutes and mount slides with aquamount I hope this helps, please let me know if you have any additional questions.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Cell lysate - nuclear (muscle)
Total protein in input
40 µg
Specification
muscle
Immuno-precipitation step
Protein A/G

Abcam user community

Verified customer

Submitted May 01 2007

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

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