Pax6 Antibody [Unconjugated]

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.

HeLa human cervical epithelial carcinoma cell line was stained with Sheep Anti-Human Pax6 Affinity-Purified Polyclonal Antibody (Catalog # AF8150, filled histogram) or Sheep IgG control Antibody (Catalog # 5-001-A, open histogram) followed by anti-Sheep IgG PE-conjugated 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.

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 the12-230 kDa separation system.

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.

• Reactivity: Hu
• Applications: WB, Simple Western, Flow, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Pax6 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Pax6
  Met1-Arg272
  Accession # P26367
• Specificity: Detects human Pax6 in direct ELISAs. In direct ELISAs, less than 5% cross-reactivity with recombinant human (rh) Pax1, rhPax2, rhPax3, rhPax4, rhPax5, and rhPax7 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: PAX6

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5 ug/mL
  • Simple Western 5 ug/mL
  • Immunocytochemistry 5-15 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Pax6 Antibody [Unconjugated]

• keratitis)
• MGC17209
• Oculorhombin
• paired box 6
• paired box protein Pax-6
• Pax6

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Pax6 Antibody (AF8150)(3)

Publications using AF8150

• A Filippello, F Urbano, S Di Mauro, A Scamporrin, A Di Pino, R Scicali, AM Rabuazzo, F Purrello, S Piro Chronic Exposure to Palmitate Impairs Insulin Signaling in an Intestinal L-cell Line: A Possible Shift from GLP-1 to Glucagon Production Int J Mol Sci, 2018;19(12):. 2018 [PMID: 30487448] (Western Blot, Mouse)
• X Gao, JJ Yourick, RL Sprando Generation of nine induced pluripotent stem cell lines as an ethnic diversity panel Stem Cell Res, 2018;31(0):193-196. 2018 [PMID: 30099336] (ICC, Human)
• WA Pastor, W Liu, D Chen, J Ho, R Kim, TJ Hunt, A Lukianchik, X Liu, JM Polo, SE Jacobsen, AT Clark TFAP2C regulates transcription in human naive pluripotency by opening enhancers Nat. Cell Biol., 2018;20(5):553-564. 2018 [PMID: 29695788] (Western Blot, Human)

FAQs for Pax6 Antibody (AF8150). (Showing 1 - 1 of 1 FAQs).

1. What PAX6 antibody is recommend to use in ICC staining of human stem cells differentiated neural precursors?
   • By performing a search on PAX6 then refining this to antibodies which have been validated with human samples and for ICC, I can see that we have three products to meet your requirements. You can see all three of these antibodies. These products are listed based on sales and citations, however all of our antibodies are covered by our 100% guarantee to work in the species and applications listed on our website and on our product datasheets. You can read more about our quality guarantee here.

Bioinformatics

• Gene Symbol: PAX6
• Uniprot:
  • P26367()