Intracellular staining of Pax-6 in neural induction of human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) were cultured in mTeSR® (Stem Cell Technologies) on plates coated with BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277). Embryoid bodies (EB) were made and cultured in medium containing Knockout™ Serum Replacement (Life Technologies) without bFGF for 24 hours and then in medium containing 250 ng/ml human recombinant noggin (R&D Systems) and 10 μM SB 431542 (Tocris) for 4 more days. The EB were then plated on BD Matrigel-coated plates and grown in medium with ITS supplement (Sigma-Aldrich), noggin, and SB 431542. After growth for 7 days, the cells were collected, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with PE Mouse anti-Human Pax-6 and PerCP-Cy5.5 Mouse anti-Human Sox1 (Cat. No. 561549). The plot was derived from gated events based on light scattering characteristics for the neural induction. Flow cytometry was performed on a BD LSR™ II flow cytometry system.
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Intracellular staining of Pax-6 in neural induction of human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) were cultured in mTeSR® (Stem Cell Technologies) on plates coated with BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277). Embryoid bodies (EB) were made and cultured in medium containing Knockout™ Serum Replacement (Life Technologies) without bFGF for 24 hours and then in medium containing 250 ng/ml human recombinant noggin (R&D Systems) and 10 μM SB 431542 (Tocris) for 4 more days. The EB were then plated on BD Matrigel-coated plates and grown in medium with ITS supplement (Sigma-Aldrich), noggin, and SB 431542. After growth for 7 days, the cells were collected, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with PE Mouse anti-Human Pax-6 and PerCP-Cy5.5 Mouse anti-Human Sox1 (Cat. No. 561549). The plot was derived from gated events based on light scattering characteristics for the neural induction. Flow cytometry was performed on a BD LSR™ II flow cytometry system.
제품 세부 정보
BD Pharmingen™
Hu PAX-6 PE O18-1330 50Tst
Oculorhombin, Aniridia type II protein, PAX6, AN2
Human (QC Testing)
Mouse BALB/c IgG2a, κ
Human Pax-6 aa 406-422 Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
5080
AB_10714781
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
준비 및 보관
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
제품 고시
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
관련 제품
Stain Buffer (FBS) RUO
사이즈
500 mL
카탈로그 No
554656
Perm Buffer III RUO
사이즈
125 mL
카탈로그 No
558050
Fixation Buffer RUO
사이즈
100 mL
카탈로그 No
554655
PE Mouse IgG2a, κ Isotype Control RUO
사이즈
50 Tests
카탈로그 No
558595
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561552 Rev. 1
항체 세부 정보
O18-1330
Pax-6 is a member of the paired box (pax) gene family whose protein products are transcription factors involved in development. Pax family members share a highly conserved DNA binding domain that contains six alpha helices (paired domain) and a homeo box domain. Pax-6 has important roles in the development of the eye, nose, central nervous system, and pancreas. Defects in Pax-6 are responsible for various eye malformations including aniridia and Peters anomaly.
The O18-1330 monoclonal antibody reacts with human Pax-6. Because the Pax-6 protein sequence is highly conserved among vertebrate species, cross-reactivity with other species is possible.
561552 Rev. 1
형광 세부 정보
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
561552 Rev.1
인용 및 참고 문헌
개발 참고 자료 (4)
-
Cerf ME. Transcription factors regulating beta-cell function. Eur J Endocrinol. 2006; 155(5):671-679. (Biology). 참조 보기
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Chambers SM, Fasano CA, Papapetrou EP, Tomishima M, Sadelain M, Studer L. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol. 2009; 27(3):275-280. (Methodology). 참조 보기
-
Glaser T, Walton DS, Maas RL. Genomic structure, evolutionary conservation and aniridia mutations in the human PAX6 gene. Nat Genet. 1992; 2:232-239. (Biology). 참조 보기
-
Osakada F, Jin ZB, Hirami Y, et al. In vitro differentiation of retinal cells from human pluripotent stem cells by small-molecule induction. J Cell Sci. 2009; 122:3169-3179. (Methodology). 참조 보기
561552 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
