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RecombinantRabMAb

Recombinant Anti-PAX6 antibody [EPR15858] (ab195045)

Reviews (8) Submit a question References (43)
Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
  • Anti-PAX6 antibody [EPR15858] (ab195045)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15858] to PAX6
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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. This information is proprietary to Abcam and/or its suppliers.

  • 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, mouse brain.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

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)
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF (5)
1/350.
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

    see all

  • 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

  • Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)
    Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)

    Ab195045 staining Pax6 in primary mouse neurons/glia, DIV1 (top row) and DIV14 (bottom row) both prepared from E18 mouse hippocampal brain area (obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP). The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab195045 at 0.2 µg/ml and ab7291, mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    A subset of cells showed staining in the nucleus at DIV1 (likely neuroprogenitor cells), while differentiated neurons (DIV14) where Pax6 negative.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)
    Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)

    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.

  • Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
    Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)
    Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)

    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.

     

  • Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
    Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)

    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.

  • Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
    Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)

    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.

  • Anti-PAX6 antibody [EPR15858] (ab195045)
    Anti-PAX6 antibody [EPR15858] (ab195045)

References (43)

Publishing research using ab195045? Please let us know so that we can cite the reference in this datasheet.

ab195045 has been referenced in 43 publications.

  • Parichha A  et al. Constitutive activation of canonical Wnt signaling disrupts choroid plexus epithelial fate. Nat Commun 13:633 (2022). PubMed: 35110543
  • Fu Y  et al. Loss of neurodevelopmental-associated miR-592 impairs neurogenesis and causes social interaction deficits. Cell Death Dis 13:292 (2022). PubMed: 35365601
  • Sun Y  et al. Involvement of Cancer Stem Cells in Chemoresistant Relapse of Epithelial Ovarian Cancer Identified by Transcriptome Analysis. J Oncol 2022:6406122 (2022). PubMed: 35401749
  • Sanaki-Matsumiya M  et al. Periodic formation of epithelial somites from human pluripotent stem cells. Nat Commun 13:2325 (2022). PubMed: 35484123
  • Brezovakova V  et al. Astrocytes Derived from Familial and Sporadic Alzheimer's Disease iPSCs Show Altered Calcium Signaling and Respond Differently to Misfolded Protein Tau. Cells 11:N/A (2022). PubMed: 35563735
View all Publications for this product

Customer reviews and Q&As

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PAX6 antibody [EPR15858]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (embryonic mouse brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 1x DAKO antigen retrieval buffer
Permeabilization
Yes - 0.3% Triton X-100 in PBS for 5 minutes
Specification
embryonic mouse brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Laura Lahti

Verified customer

Submitted Dec 03 2021

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