GloLIVE Human/Mouse SSEA-1 NL557 Live Cell Antibody

R&D Systems | Catalog # NLLC2155R

50X Concentration, Azide Free
R&D Systems
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Key Product Details

Species

Human, Mouse

Applications

Immunocytochemistry/ Immunofluorescence

Conjugate

NL557

Product Summary for GloLIVE Human/Mouse SSEA-1 NL557 Live Cell Antibody

Kit Summary

To detect SSEA-1 expression in live cells by immunocytochemical staining.

Key Benefits

  • Verifies marker expression in live stem cells
  • Requires only one 30 minute incubation
  • Permits continued cell culture post-staining
  • Does not affect cell proliferation or stemness

NLLC2155R SSEA-1 Example Data
View Data

 

 

Why confirm marker expression in live cells before colony selection?

Maintaining pluripotent human embryonic stem cells and deriving induced pluripotent stem cells require colony selection and cell expansion. Picking colonies that contain pluripotent stem cells is typically achieved by manually analyzing colony morphology. Unfortunately, this critical process is time-consuming and labor-intensive.

In addition, morphology-based colony selection does not consistently guarantee pluripotent starting populations, since cells can fail to be fully reprogrammed or can naturally differentiate within a colony.

To address this need, R&D Systems offers GloLIVE™ antibodies that allow researchers to confirm stem cell marker expression in live cells before colony selection.

Live pluripotent stem cell imaging:

  • Promotes the selection of high quality, undifferentiated stem cell colonies to reduce experimental variation.
  • Verifies stem cell pluripotency in 30 minutes, ensuring efficient use of time and reagents.
  • Allows for the continued culture of pluripotent cells with no adverse effects on cell proliferation or stemness following staining.

 

Antibody Components
  • GloLIVE NL557-conjugated Mouse Anti-Human/Mouse SSEA-1 Monoclonal Antibody.
  • Supplied as 50X concentration of antibody in 0.5 mL PBS

Azide free and endotoxin tested (= 5 EU/mL), this antibody can be used for 12 tests, if a staining volume of 2 mL is used.

 

Data
Detection of SSEA-1 in Live D3 Mouse Embryonic Stem Cells
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Detection of SSEA-1 in Live D3 Mouse Embryonic Stem Cells. The stem cell marker SSEA-1 was visualized in live D3 mouse embryonic stem cells, grown on irradiated mouse embryonic fibroblasts (Catalog # PSC001), using a GloLIVE NL557-conjugated Anti-Human/Mouse SSEA-1 Monoclonal Antibody (Catalog # NLLC2155R; red). The nuclei were counterstained with Hoechst 33342 (blue). Positive staining for SSEA-1 in mouse stem cells is an indicator of pluripotency.

Product Specifications

Specificity

Detects human and mouse SSEA-1.

Clone

MC-480

Scientific Data Images for GloLIVE Human/Mouse SSEA-1 NL557 Live Cell Antibody

Detection of SSEA-1 in Live D3 Mouse Embryonic Stem Cells.

The stem cell marker SSEA-1 was visualized in live D3 mouse embryonic stem cells, grown on irradiated mouse embryonic fibroblasts (Catalog # PSC001), using a GloLIVE NL557-conjugated Anti-Human/Mouse SSEA-1 Monoclonal Antibody (Catalog # NLLC2155R; red). The nuclei were counterstained with Hoechst 33342 (blue). Positive staining for SSEA-1 in mouse stem cells is an indicator of pluripotency.

Formulation, Preparation, and Storage

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store the unopened product at 2 - 8° C. Do not use past expiration date. Protect from light.

Background: SSEA-1

Stage-Specific Embryonic Antigen-1 (SSEA-1), an antigenic epitope defined as a Lewis x carbohydrate structure, is expressed during early mouse embryogenesis on murine embryonal carcinoma cells (EC), murine embryonic stem cells (ES), and murine and human germ cells. Expression of SSEA-1 is down-regulated following differentiation of murine EC and ES cells. In contrast, the differentiation of human EC and ES cells is accompanied by an increase in SSEA-1 expression (1, 2).

References

  1. Solter, D. and Knowles, B.B. (1978) Proc. Natl. Acad. Sci. USA 75:5565.
  2. Fox, N. et al. (1983) Cancer Res. 43:669.

Long Name

Stage-specific Embryonic Antigen-1

Alternate Names

SSEA1

Additional SSEA-1 Products

Product Documents for GloLIVE Human/Mouse SSEA-1 NL557 Live Cell Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for GloLIVE Human/Mouse SSEA-1 NL557 Live Cell Antibody

For research use only

Citations for GloLIVE Human/Mouse SSEA-1 NL557 Live Cell Antibody

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Protocols

View specific protocols for GloLIVE Human/Mouse SSEA-1 NL557 Live Cell Antibody (NLLC2155R):

Refer to the product datasheet for complete product details.

Briefly, SSEA-1 expression can be verified in live cells prior to colony selection or experimentation using this simple procedure:
  • Add the SSEA-1 antibody directly to the media of cultured cells
  • Wash the cells after a 30 minute incubation
  • Select for and expand SSEA-1-positive colonies

 
 

Precautions

Use aseptic technique and sterile culturing conditions to prevent contamination.

Reagents Provided

GloLIVE NL557-conjugated Mouse Anti-Human/Mouse SSEA-1 Monoclonal Antibody. (Catalog # NLLC2155R):

Other Supplies Required

Reagents

  • Stem cell culture media

Materials

  • Human or mouse pluripotent stem cells
  • Pipettes and pipette tips
  • Serological pipette

Equipment

  • 37 °C and 5% CO2 incubator
  • Fluorescence microscope
 

Live Cell Immunocytochemistry Procedure Overview

To ensure sterility of cultures, all steps should be performed under sterile conditions.

Add primary antibody in appropriate culture media to cells.
 
  1. Add fluorochrome-conjugated primary antibody in appropriate culture media to cells.
  2. Incubate for 30 minutes.
Replace with fresh culture media
 
  1. Replace with fresh culture media.
Visualize using a fluorescence microscope
 
  1. Visualize using a fluorescence microscope.

Note: Culture with GloLIVE antibodies does not appear to affect cell proliferation or stemness as assayed by proliferation curves for 3 days post-antibody incubation and expression levels of SSEA-1 and Oct-3/4.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for GloLIVE Human/Mouse SSEA-1 NL557 Live Cell Antibody

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  • Q: What is the recommended starting confluence to observe staining?

    A: The optimal confluence would depend on the purpose of the staining. If the plan is to use all the cells in the flask after they have proliferated, then one can wait for the cells to become confluent or almost confluent (80-90% confluent) and then check the staining. If the goal is to to check on the cells mid-way through their growth and there are sufficient cells in the flask, one could check the staining then as well. The staining is not dependent on how many cells are in the flask, as long as there are sufficient cells.

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