Overview

  • Product name

    Anti-PAX6 antibody [EPR15858]
    See all PAX6 primary antibodies
  • Description

    Rabbit monoclonal [EPR15858] to PAX6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human PAX6 aa 150-350. The exact sequence is proprietary.
    Database link: P26367

  • Positive control

    • IHC-P: Mouse eyeball,Human pancreas, cerebellum, retina and retinoblastoma tissues WB: Y79 and HeLa cell lysates; PAX6 transfected 293T cell lysate. ICC/IF: Y79 and Neuro-2a cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab195045 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 1/1000. Detects a band of approximately 47, 33, 32 kDa (predicted molecular weight: 47 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/350.

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

  • Immunofluorescence analysis of 4% paraformaldehyde-fixed, 0.1% tritonX-100 Neuro-2a (mouse neuroblastoma) cells labeling PAX6 with ab195045 at 1/350, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 (green).

    Confocal image showing cytoplasmic and nuclear staining on Neuro-2a cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 (red).  

    The negative controls are as follows:-

    -ve control 1: ab195045 at 1/350 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.

    -ve control 2: ab7291 at 1/500 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400.

  • Immunohistochemical analysis of paraffin-embedded Human retina tissue labeling PAX6 with ab195045 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Nuclear staining on human retina tissue is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-PAX6 antibody [EPR15858] (ab195045) at 1/10000 dilution

    Lane 1 : PAX6 transfected 293T cell lysate
    Lane 2 : Non-transfected 293T cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 32,33,47 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    The Transfected lysate spans the immunogenic region: aa1-aa422(47kDa).

  • Immunofluorescence analysis of 4% paraformaldehyde-fixed, 0.1% tritonX-100 Y79 (Human retinoblastoma cell line) cells labeling PAX6 with ab195045 at 1/350, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 (green).

    Confocal image showing cytoplasmic and nuclear staining on Y79 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 (red).  

    The negative controls are as follows:-
    -ve control 1: ab195045 at 1/350 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.

    -ve control 2: ab7291 at 1/500 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400.

  • IHC image of Pax6 staining in a formalin fixed, paraffin embedded normal human cerebellum 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 ab195045, 1/1000 dilution, 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.

     

  • Anti-PAX6 antibody [EPR15858] (ab195045) at 1/1000 dilution + Mouse eyeball lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 32,33,47 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    The 47kDa band represents the full length PAX6, we hypothesis the 32kDa & 33kDa bands represent the PAX6p32 & PAX6p33 fragments.

  • Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling PAX6 with ab195045 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Nuclear staining on human pancreas tissue is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-PAX6 antibody [EPR15858] (ab195045) at 1/10000 dilution

    Lane 1 : Y79 cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 32,33,47 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    The 47kDa band represents the full length PAX6, we hypothesis the 32kDa & 33kDa bands represent the PAX6p32 & PAX6p33 fragments.

  • Immunohistochemical analysis of paraffin-embedded Human retinoblastoma tissue labeling PAX6 with ab195045 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Nuclear staining on human retinoblastoma tissue is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • IHC image of Pax6 staining in a formalin fixed, paraffin embedded normal rat cerebellum 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 ab195045, 1/1000 dilution, 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.

References

This product has been referenced in:

  • Boisvert EM  et al. Minocycline mitigates the effect of neonatal hypoxic insult on human brain organoids. Cell Death Dis 10:325 (2019). Read more (PubMed: 30975982) »
  • Bai X  et al. Generation of an integration-free induced pluripotent stem cell line (FDEENTi003-A) from a patient with pathological myopia. Stem Cell Res 39:101495 (2019). Read more (PubMed: 31376721) »
See all 4 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cultured Cells (iPSC-derived neuron progenitor)
Permeabilization
Yes - 0.1% Triton X-100
Specification
iPSC-derived neuron progenitor
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 01 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Pancreas)
Permeabilization
No
Specification
Pancreas
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 20 2019

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (adult mouse brain, lateral ventricle/SVZ)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization
Yes - 1% Triton X in PBS
Specification
adult mouse brain, lateral ventricle/SVZ
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10 · Temperature: 21°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 15 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Differentiated Embryonic Stem Cells)
Permeabilization
Yes - 0.5% Triton X-100
Specification
Differentiated Embryonic Stem Cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Mr. Dylan Stavish

Verified customer

Submitted May 01 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (3D neuronal tissue)
Permeabilization
No
Specification
3D neuronal tissue
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Formaldehyde

Mr. Tamas Bellak

Verified customer

Submitted Apr 24 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Differentiated Human iPS cells)
Permeabilization
Yes - 0.1% triton X
Specification
Differentiated Human iPS cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 21 2016

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