Overview

  • Product name

    Anti-PAX6 antibody [AD2.38]
    See all PAX6 primary antibodies
  • Description

    Mouse monoclonal [AD2.38] to PAX6
  • Host species

    Mouse
  • Specificity

    This antibody recognizes both products of the two major alternatively spliced forms. In our hands this product does not detect a band of interest in Western Blot.
  • Tested applications

    Suitable for: IHC-Fr, Sandwich ELISA, IHC-P, ICC/IF, IHC-FoFrmore details
    Unsuitable for: WB
  • Species reactivity

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

    Recombinant full length protein corresponding to Human PAX6.

  • Positive control

    • IHC-P: FFPE Human pancreas normal, Rat retina.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab78545 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
IHC-Fr Use at an assay dependent concentration.
Sandwich ELISA Use a concentration of 5 µg/ml. For sandwich ELISA, use this antibody as Capture at 5 µg/ml with Rabbit polyclonal to PAX6 (ab82510) as Detection.
IHC-P Use a concentration of 10 - 20 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for WB.
  • Target

    • Function

      Transcription factor with important functions in the development of the eye, nose, central nervous system and pancreas. Required for the differentiation of pancreatic islet alpha cells (By similarity). Competes with PAX4 in binding to a common element in the glucagon, insulin and somatostatin promoters. Regulates specification of the ventral neuron subtypes by establishing the correct progenitor domains (By similarity). Isoform 5a appears to function as a molecular switch that specifies target genes.
    • Tissue specificity

      Fetal eye, brain, spinal cord and olfactory epithelium. Isoform 5a is less abundant than the PAX6 shorter form.
    • Involvement in disease

      Defects in PAX6 are the cause of aniridia (AN) [MIM:106210]. A congenital, bilateral, panocular disorder characterized by complete absence of the iris or extreme iris hypoplasia. Aniridia is not just an isolated defect in iris development but it is associated with macular and optic nerve hypoplasia, cataract, corneal changes, nystagmus. Visual acuity is generally low but is unrelated to the degree of iris hypoplasia. Glaucoma is a secondary problem causing additional visual loss over time.
      Defects in PAX6 are a cause of Peters anomaly (PAN) [MIM:604229]. Peters anomaly consists of a central corneal leukoma, absence of the posterior corneal stroma and Descemet membrane, and a variable degree of iris and lenticular attachments to the central aspect of the posterior cornea.
      Defects in PAX6 are a cause of foveal hypoplasia (FOVHYP) [MIM:136520]. Foveal hypoplasia can be isolated or associated with presenile cataract. Inheritance is autosomal dominant.
      Defects in PAX6 are a cause of keratitis hereditary (KERH) [MIM:148190]. An ocular disorder characterized by corneal opacification, recurrent stromal keratitis and vascularization.
      Defects in PAX6 are a cause of coloboma ocular (COLO) [MIM:120200]; also known as uveoretinal coloboma or coloboma of iris, choroid and retina. Ocular colobomas are a set of malformations resulting from abnormal morphogenesis of the optic cup and stalk, and the fusion of the fetal fissure (optic fissure). Severe colobomatous malformations may cause as much as 10% of the childhood blindness. The clinical presentation of ocular coloboma is variable. Some individuals may present with minimal defects in the anterior iris leaf without other ocular defects. More complex malformations create a combination of iris, uveoretinal and/or optic nerve defects without or with microphthalmia or even anophthalmia.
      Defects in PAX6 are a cause of coloboma of optic nerve (COLON) [MIM:120430].
      Defects in PAX6 are a cause of bilateral optic nerve hypoplasia (BONH) [MIM:165550]; also known as bilateral optic nerve aplasia. A congenital anomaly in which the optic disc appears abnormally small. It may be an isolated finding or part of a spectrum of anatomic and functional abnormalities that includes partial or complete agenesis of the septum pellucidum, other midline brain defects, cerebral anomalies, pituitary dysfunction, and structural abnormalities of the pituitary.
      Defects in PAX6 are a cause of aniridia cerebellar ataxia and mental deficiency (ACAMD) [MIM:206700]; also known as Gillespie syndrome. A rare condition consisting of partial rudimentary iris, cerebellar impairment of the ability to perform coordinated voluntary movements, and mental retardation.
    • Sequence similarities

      Belongs to the paired homeobox family.
      Contains 1 homeobox DNA-binding domain.
      Contains 1 paired domain.
    • Developmental stage

      Expressed in the developing eye and brain.
    • Post-translational
      modifications

      Ubiquitinated by TRIM11, leading to ubiquitination and proteasomal degradation.
    • Cellular localization

      Nucleus.
    • Information by UniProt
    • Database links

    • Alternative names

      • AN 2 antibody
      • AN antibody
      • AN2 antibody
      • Aniridia type II protein antibody
      • D11S812E antibody
      • FVH1 antibody
      • KIAA0552 antibody
      • Leucine zipper putative tumor suppressor 3 antibody
      • LZTS3 antibody
      • MGC17209 antibody
      • MGDA antibody
      • Oculorhombin antibody
      • Paired box 6 antibody
      • Paired box gene 6 (aniridia keratitis) antibody
      • Paired Box Gene 6 antibody
      • Paired box homeotic gene 6 antibody
      • Paired box protein Pax-6 antibody
      • Paired box protein Pax6 antibody
      • PAX 6 antibody
      • PAX6 antibody
      • PAX6_HUMAN antibody
      • ProSAP-interacting protein 1 antibody
      • PROSAPIP1 antibody
      • Sey antibody
      • WAGR antibody
      see all

    Images

    • IHC image of Pax6 staining in a formalin fixed, paraffin embedded normal rat retina tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab78545, 5µg/ml, for 15 mins 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.

       

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • IHC image of PAX6 staining in a formalin fixed, paraffin embedded normal human pancreas tissue section*performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab78545, 20µg/ml, for 15 mins 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.

       For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Immunohistochemical analysis of PFA-fixed frozen murine embryonic brain coronal sections, labelling PAX6 with ab78545 at a dilution of 1/100 incubated for 8 hours at 4°C in blocking buffer diluent. Permeabilization was with Triton X-100 and blocking was with 1% serum for 1 hour. Heat mediated antigen retrival was with 10mM sodium citrate buffer pH6.0 for 10 minutes at 650W in a microwave. The secondary was a goat Alexa Fluor® 488 at 1/700.

      See Abreview

    • IHC image of PAX6 staining in mouse e14 foetus formalin fixed paraffin embedded tissue section, with the use of Mouse on Mouse Polymer IHC Kit (Ab127055). The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab78545, 10µg/ml overnight at +4°C. The Mouse on Mouse HRP Polymer was incubated for 15 minutes at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

    • Standard Curve for Pax6; dilution range 1 pg/ml to 1 ug/ml using Capture Antibody Mouse monoclonal [AD2.38] to PAX6 (ab78545) at 5 ug/ml and Detector Antibody Rabbit polyclonal to PAX6 (ab82510) at 0.5 ug/ml.
    • Immunohistochemical analysis of mouse brain tissue, staining PAX6 with ab78545.

      Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in PBST) for 12 hours at 4°C. An AlexaFluor®568-conjugated goat anti-mouse polyclonal IgG (1/1000) was used as the secondary antibody.

      See Abreview

    References

    This product has been referenced in:

    See all 21 Publications for this product

    Customer reviews and Q&As

    1-10 of 17 Abreviews or Q&A

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (brain organoid)
    Permeabilization
    Yes - PBS-Triton 0.5%
    Specification
    brain organoid
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Sep 11 2017

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (embryonic mouse brain coronal tissue section)
    Permeabilization
    Yes - Triton-X 100
    Specification
    embryonic mouse brain coronal tissue section
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
    Fixative
    Paraformaldehyde

    Dr. Bhavin Shah

    Verified customer

    Submitted Oct 19 2015

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (brainstem)
    Permeabilization
    Yes - 0.1% triton
    Specification
    brainstem
    Blocking step
    BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 10 2015

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (E15.5 brain)
    Specification
    E15.5 brain
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Nov 05 2012

    Question

    Hello,
    Here are technical details which you asked for.
    Sincerely,

    General Information Antibody storage conditions (temperature/reconstitution etc) room temp for 48 hours, afterwards +4C

    Description of the problem (high background, low signal, non-specific staining etc.) - no nuclear staining, very weak cytoplasmic staining (for PAX6 nuclear staining is of central importance)

    Sample (Species/Tissue/Cell Type/Cell Line etc.) HPAF-II cells for ICC, pancreatic adenocarcinoma with normal areas for IHC

    Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.) ICC - 4% paraformaldehyde, for IHP - standard formalin fixation

    Antigen retrieval (Enzymatic method, Heat mediated technique etc.) citrate pH 6 and EDTA-Tris pH 9

    Permeabilization step - - methanol and acetone tested for ICC , 0,25% Triton-X100 for IHC

    Blocking conditions (Buffer/time period, Blocking agent etc.) - 3% rabbit serum

    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) antibody in question was diluted 1:100, 1:200, 1:500, 1:1000, incubation 1h at room temp, washes performed with 1xPBS

    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) - DAKO a-mouse kit for IHC, anti-mouse Alexa 555 1:500 for ICC

    Detection method - DAB for IHC, confocal microscopy for ICC

    Positive and negative controls used (please specify) - pancreas is itself a positive control for PAX6

    Optimization attempts (problem solving) How many times have you tried the IHC? - 4 dilutions in 2 antigen retrieval protocols

    Have you run a No Primary control? - Yes

    Do you obtain the same results every time? - Yes

    Read More
    Answer

    Thank you for taking the time to complete our questionnaire.

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    Reviewing this case, I would like to offer some suggestions to help optimise the results from ab78545. I would also appreciate if you can confirm some further details:

    1. I can recommend to aliquot and store at -20oC as directed on the datasheet in order to ensure the antibody remains stable and we can continue to provide the guarantee.

    2. I can recommend to consider washing with PBS containing 0.2% Tween, a more gentle detergent, also to add 0.2% Tween to the antibody dilution buffer. This will help keep the antibody solubilised and keep the cells permeabilized.

    3. The time for antigen retrieval can sometimes require some optimization. I can suggest to try 2, 5 10 and 20 minutes if this has not already been tried.

    4. I would appreciate if you are able to provide an image which would help me to assess the results.

    5. Try longer incubation, overnight at 4oC. This can sometimes provide more efficient and specific staining.

    6. I can suggest permeabilization should be for 10 minutes only with these reagents if this has not already been tried. Otherwize the staining could be affected. As this is a nuclear target, try permeabilizing the IHC-P samples too.

    7. Fixation in ICC is usually 10 minutes only if this has not already been tried.

    8. Is the secondary antibody and detection kit working well with other primary antibodies? What were the results of the no primary control?

    I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More

    Answer

    Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

    I would like to reassure you that this antibody is tested and covered by our guarantee for Mouse, Rat, Chicken, and Human samples, and for IHC-P and ICC/IF. Data we have indicates that warming to room temperature over a weekend should not significantly affect the performance of the antibody, though of course we will still take this into consideration. Although the other antibody has worked well, individual antibodies can often require individual optimimization. Therefore, before deciding how to proceed, I would like to investigate this particular case further for you and obtain some further information that will be beneficial for our quality monitoring records.

    In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

    Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.


    Order Details Antibody code:
    Lot number:
    Purchase order number or preferably Abcam order number:
    General Information Antibody storage conditions (temperature/reconstitution etc)
    Description of the problem (high background, low signal, non-specific staining etc.)
    Sample (Species/Tissue/Cell Type/Cell Line etc.)
    Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)
    Antigen retrieval (Enzymatic method, Heat mediated technique etc.)
    Permeabilization step
    Blocking conditions (Buffer/time period, Blocking agent etc.)
    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
    Detection method
    Positive and negative controls used (please specify)
    Optimization attempts (problem solving) How many times have you tried the IHC?
    Have you run a "No Primary" control? Yes No
    Do you obtain the same results every time? Yes No
    What steps have you altered?
    Additional Notes
    We would appreciate if you are also able to provide and image which would help us to assess the results

    Read More

    Answer

    Thank you for your reply.

    Your credit note ID is **.

    I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

    The credit note ID is for your reference only and does not automatically guarantee the credit.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

    Read More

    Answer

    Thank you for your reply.

    I am CC'ing your lab manager on this reply as requested.

    If you could provide the order number or PO number that was used to purchase ab78545, then I will go ahead and refund the cost of the antibody to you.

    I look forward to your reply.

    Read More

    Answer

    Thank you for providing that image.

    The difference in staining between the Pax6 and Tbr2 makes me think that protocol tips would not be effective in resolving this staining issue. Therefore I would like to either replace or refund the antibody. A suitable alternative would be, ab5790:

    https://www.abcam.com/PAX6-antibody-Stem-Cell-Marker-ab5790.html



    Please let me know how you would like to proceed and if you could also include the order number or PO number that was used to purchase ab78545, then that would be very helpful.

    I look forward to your reply.

    Read More

    Question
    Answer

    Thank you for calling Abcam earlier today.

    I was wandering if you would be able to send an image of the staining that you are getting when you stain you mouse tissue with ab78545.

    I would just like to assure you that the antibody is covered under our Abpromise for six months and is guaranteed to work in IHC-P on mouse samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

    I look forward to your reply and helping resolve this issue.

    Read More

    1-10 of 17 Abreviews or Q&A

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