Invitrogen

SSEA4 Monoclonal Antibody (MC-813-70), DyLight™ 488

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SSEA4 Antibody in Immunocytochemistry (ICC/IF) — Imunofluorescent analysis of SSEA4 (green) in human episomal iPSC cells. Cells were grown on vitronectin coated plates, fixed with 4% paraformaldehyde for 15 minutes, and blocked with 3% Blocker BSA (Product # 37525) for 30 minutes at room temperature. Cells were stained with DyLight 488-conjugated SSEA4 monoclonal antibody (left panels, Product # MA1-021-D488) or DyLight 488-conjugated Mouse IgG3 isotype control antibody (top right panel, Product # MA1-190-D488), at a dilution of 1:50 and incubated for 1 hour at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on an EVOS FL Imaging system at 20X magnification.

SSEA4 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of SSEA-4 in NCCIT (green filled histogram) and HeLa cells (green unfilled histogram) compared to unstained cells (blue histogram). Positive staining is observed on NCCIT cells when compared to no antibody control (middle panel) and to SSEA-4-negative HeLa cells (right panel). Cells were fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with 2% BSA-PBS for 30 minutes at room temperature and incubated with a DyLight 488-conjugated SSEA-4 monoclonal antibody (Product # MA1-021-D488) in 2% BSA-PBS at a dilution of 1:100 for 60 minutes at room temperature. Cells were washed and re-suspended in PBS for FACS analysis.

SSEA4 Antibody in Immunohistochemistry (IHC) — Figure 5 FACT Depletion Increases Efficiency of Reprogramming Human Fibroblasts (A) Reprogramming with human secondary fibroblasts carrying a doxycycline (DOX)-inducible OCT4/SOX2/KLF4/c-MYC (OSKM) cassette ( Cacchiarelli et al., 2015 ) that were transfected with siRNAs against human SSRP1 or SUPTH16 before DOX induction. (B) Alkaline phosphatase (AP) staining of stem cell colonies 21 days after DOX treatment. Control experiment is scrambled siRNA (see STAR Methods ). (C) Quantification of iPSC colonies (6 replicates) based on SSEA4 immunostaining for each knockdown condition. Paired Student's t test was used for statistical comparison, * p < 0.05, *** p < 0.001. Error bars represent SD. (D) Histological characterization of teratomas derived from grafting SSRP1 or SUPT16H depletion-derived iPSCs in mice. Teratomas reached 1.5 cm 2 in size after 51-70 days and showed tissues of all three germ layers (ectoderm, endoderm, mesoderm). Scale bars represent 50 mum. (E) Forced expression of Ascl1, Brn2, and Mytl1 in human fibroblasts (Normal Human Dermal Fibroblats = NHDF cells) and SSRP1 or SUPT16H depletion enhances reprogramming of fibroblasts to neurons. One-way ANOVA test was used for statistical comparison, ** p < 0.01. Error bars represent SEM. See also Figure S5 .

产品信息

MA1-021-D488

应用	建议稀释比	已发表文章
免疫组化 (IHC)	-	查看1篇已发表文章 1篇已发表文章
免疫细胞化学 (ICC/IF)	1:50-1:500	-
流式细胞分析 (Flow)	1:50-1:500	-

产品规格
种属反应	Human
已发表种属	Not Applicable
宿主/亚型	Mouse / IgG3
分类	Monoclonal
类型	Antibody
克隆号	MC-813-70
抗原	human embryonal carcinoma cell line 2102Ep
偶联物	DyLight™ 488 Alexa Fluor 647 DyLight 550 DyLight 650 PE Unconjugated
激发/发射光谱	查看光谱
形式	Liquid
浓度	1 mg/mL
纯化类型	Protein A
保存液	PBS with proprietary stabilizer
内含物	0.02% sodium azide
保存条件	4° C, do not freeze
RRID	AB_2536688

产品详细信息

MA1-021-D488 has been successfully used in ICC/IF and flow cytometry applications on human samples.

靶标信息

SSEA-4 (Stage-specific embryonic antigen 4) is a cell surface antigen, expressed along with SSEA-3, TRA-1-60 and TRA-1-81 in embryonic stem cells, embryonal carcinoma cells and induced pluripotent stem cells (iPS). In human cells, these surface markers are down-regulated during the differentiation process, while SSEA-1 is absent in undifferentiated human stem cells and upregulated on the cell surface after retinoic acid mediated differentiation. Conversely, in mouse stem cells, SSEA-1 expression decreases with differentiation while SSEA-3 and SSEA-4 expression increase.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

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Product References

FACT Sets a Barrier for Cell Fate Reprogramming in Caenorhabditis elegans and Human Cells.

Developmental cell

Kolundzic E,Ofenbauer A,Bulut SI,Uyar B,Baytek G,Sommermeier A,Seelk S,He M,Hirsekorn A,Vucicevic D,Akalin A,Diecke S,Lacadie SA,Tursun B

Published figure using SSEA4 monoclonal antibody (Product # MA1-021-D488) in Immunofluorescence

Mon Sep 10 00:00:00 EDT 2018

生物信息学

蛋白别名: SSEA-4; stage-specific embryonic antigen 4

SSEA4 Monoclonal Antibody (MC-813-70), DyLight™ 488

产品信息

MA1-021-D488

免疫组化 (IHC)

免疫细胞化学 (ICC/IF)

流式细胞分析 (Flow)

种属反应

已发表种属

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偶联物

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纯化类型

保存液

内含物

保存条件

RRID

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