Product Details

Reactivity

Human

Antibody Type

Monoclonal

Host Species

Mouse

Formulation

0.1 ?m filtered in phosphate-buffered solution, pH 7.2, containing no preservative. Endotoxin level is < 0.01 EU/?g of the protein (< 0.001 ng/?g of the protein) as determined by the LAL test.

Preparation

The Ultra-LEAF? (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.

Concentration

1.0 mg/mL

Storage & Handling

The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF? solution contains no preservative; handle under aseptic conditions.

Application

Activ
FC
Cell Culture

Recommended Usage

1 ?g/mL of antibody antibody in PBS, for plate coating protocols; 10 ?g/mL of OKT3 antibody for soluble antibody activation methods

Application Notes

The OKT3 monoclonal antibody reacts with an epitope on the epsilon-subunit within the human CD3 complex.

Clone OKT3 can block the binding of clones SK7 and UCHT1.⁴ The OKT3 antibody is able to induce T cell activation. Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections and activation of T cells. The LEAF? purified antibody (Endotoxin <0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 317304). For highly sensitive assays, we recommend Ultra-LEAF? purified antibody (Cat. No. 317326) with a lower endotoxin limit than standard LEAF? purified antibodies (Endotoxin <0.01 EU/?g).

Additional Product Notes

This antibody is intended for activation and expansion of CD3 T cells in research and ex vivo cell culture applications. The antibody was GMP manufactured under serum-free conditions, including serum-free hybridoma cell culture, without additional animal or human-derived materials.

OKT3 antibody can be used in combination with anti-human CD28 antibodies to efficiently activate T cells when used at 1 ?g/mL of antibody in PBS, for plate coating protocols. Plates should be coated for 2 hours in a tissue culture incubator, or overnight at 4°C. For soluble antibody activation protocols we recommend 10 ?g/mL of OKT3 antibody and 5 ?g/mL of anti-human CD28.

Application References

(PubMed link indicates BioLegend citation)

1. Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York.
2. Knapp W. 1989. Leucocyte Typing IV. Oxford University Press New York.
3. Barclay N, et al. 1997. The Leucocyte Antigen Facts Book. Academic Press Inc. San Diego.
4. Li B, et al. 2005. Immunology 116:487.
5. Jeong HY, et al. 2008. J. Leuckocyte Biol. 83:755. PubMed
6. Alter G, et al. 2008. J. Virol. 82:9668. PubMed
7. Manevich-Mendelson E, et al. 2009. Blood 114:2344. PubMed
8. Pinto JP, et al. 2010. Immunology. 130:217. PubMed
9. Biggs MJ, et al. 2011. J. R. Soc. Interface. 8:1462. PubMed

RRID

AB_2894461 (BioLegend Cat. No. 317353) AB_2894461 (BioLegend Cat. No. 317354)

Disclaimer

GMP Ultra-LEAF? antibodies. BioLegend GMP Ultra-LEAF? antibodies are manufactured in a dedicated GMP facility and compliant with ISO 13485:2016. For research or ex vivo cell processing use. Not for use in diagnostic or therapeutic procedures.　Our processes include:

• Batch-to-batch consistency　 　　
• Material traceability
• Documented procedures
• Documented employee training
• Equipment maintenance and monitoring records
• Lot-specific certificates of analysis
• Quality audits per ISO 13485:2016
• QA review of released products

BioLegend GMP Ultra-LEAF? antibodies are manufactured and tested in accordance with USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Ph. Eur. Chapter 5.2.12