Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all Pax6 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Pax6
Met1-Arg272
Accession # P26367

Specificity

Detects human Pax6 in direct ELISAs.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human Pax6 Antibody

Detection of Human and Rat Pax6 antibody by Western Blot.

Detection of Human and Rat Pax6 by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and rat cortical stem cells. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Pax6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8150) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Pax6 at approximately 48-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Pax6 antibody in SA01 Human Embryonic Stem Cells by Immunocytochemistry (ICC).

Pax6 in SA01 Human Embryonic Stem Cells.

Immersion fixed SA01 human embryonic stem cells were differentiated for 6 days with Recombinant Human Noggin (Catalog # 6057-NG) and SB431542 (upper panel) or differentiated for 6 days with Recombinant Human BMP-4 (negative control, lower panel; Catalog # 314-BP). Pax6 was detected using Sheep Anti-Human Pax6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8150) at 5 µg/mL. Cells were stained using an Alexa Fluor 488-conjugated Anti-Sheep IgG Secondary Antibody (green) and counterstained with DAPI (blue). Specific staining was localized to nuclei.Images courtesy of Dr. Ron McKay, Leiber Institute for Brain Development, Baltimore, Maryland, USA.

Detection of Human Pax6 antibody by Simple WesternTM.

Detection of Human Pax6 by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Pax6 at approximately 59 kDa (as indicated) using 5 µg/mL of Sheep Anti-Human Pax6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8150) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Pax6 in Jurkat cells by Flow Cytometry.

Jurkat cells were stained with Sheep Anti-Human Pax6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8150, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Phycoerythrin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0126). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

Detection of Pax6 by Flow Cytometry

Detection of Pax6 by Flow Cytometry

BAF250a was dispensable for neuroectoderm differentiation. (A) FACS analysis of differentiation of WT and BAF250a KO hESC to neuroectoderm using PAX6 antibody at differentiation day 12. (B) Relative mRNA expression of TFAP2A and PAX6 at day 12 differentiation. (C) Immunoprecipitation of day 2 differentiation cells after LDN193189 and SB43142 treatment using anti-OCT4 antibody, followed by Western blotting to detect beta -CATENIN and active beta -CATENIN. Five percent of cell lysates were used as input controls. (D) Relative band intensity of precipitated beta -CATENIN and active beta -CATENIN in WT and BAF250 KO cells. ChIP assays with (E) anti-OCT4 and (F) anti-beta -CATENIN antibodies using day 2 differentiated cells after LDN193189 and SB43142 treatment at TFAP2A and PAX6 promoters. n = 3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32039194), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Pax6 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed SA01 human embryonic stem cells

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Jurkat human acute T cell leukemia cell line

Simple Western

5 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Western Blot

0.5 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and rat cortical stem cells

Reviewed Applications

Read 3 reviews rated 4.3 using AF8150 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Pax6

Pax6 (paired box 6; also Oculorhombin) is a 48‑50 kDa member of the paired homeobox family of transcription factors.  It is expressed in developing optic vesicle, olfactory dopaminergic neurons, and pancreatic endocrine cells.  Pax6 is a transactivating protein that interacts with MAF, CDX2 and SOX2.  Human Pax6 is 422 amino acids (aa) in length.  It contains an N-terminal paired box DNA-binding domain (aa 4‑130), a Gly-rich central region (aa 131‑209), a homeodomain (aa 213‑269) and a C-terminal Pro/Ser/Thr-rich regulatory domain (aa 279‑422).  Phosphorylation of the C‑terminal domain at Thr281/304/373 promotes Pax6 activity.  Multiple splice forms of Pax6 exist. There are alternative start sites at Met137 and a position 34 aa upstream of the standard site. There is also a deletion of aa 22‑26 and 37‑39, plus a 14 aa insertion after Gln47 that generates a C‑terminal DNA binding site.  Human and mouse Pax6 are absolutely identical in aa sequence.

Long Name

Paired Box Gene 6

Alternate Names

keratitis), MGC17209, Oculorhombin, paired box 6, paired box protein Pax-6

Entrez Gene IDs

5080 (Human); 18508 (Mouse); 25509 (Rat)

Gene Symbol

PAX6

UniProt

P26367

Additional Pax6 Products

Product Documents for Human Pax6 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Pax6 Antibody

For research use only

Citations for Human Pax6 Antibody

Customer Reviews for Human Pax6 Antibody (3)

4.3 out of 5
3 Customer Ratings
5 Stars
33%
4 Stars
67%
3 Stars
0%
2 Stars
0%
1 Stars
0%

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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • Human Pax6 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Neural progenitor cells
    Species: Human
    Verified Customer | Posted 09/26/2018
    Pax6 was detected human embryonic stem cells-derived neural stem cells using Sheep Anti-Human PAX6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8150) at 1 µg/mL for overnight at 4°C. Cells were stained using the Donkey anti-sheep IgG (H+L) Cross-Adsorbed, Alexa Fluor® 568.
    Human Pax6 Antibody AF8150
  • Human Pax6 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Cerebellum tissue
    Species: Mouse
    Verified Customer | Posted 11/02/2017
    Works fine at 1/1000 dilution. O.N. incubation at 4C.
    Human Pax6 Antibody AF8150
  • Human Pax6 Antibody
    Name: Juan Zanin
    Application: Immunohistochemistry
    Sample Tested: Cerebellum tissue
    Species: Mouse
    Verified Customer | Posted 07/13/2016
    Human Pax6 Antibody AF8150

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Protocols

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