Whitlow/218 Linker (E3U7Q) F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) (#47375) Datasheet with Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:

FC-L

REACTIVITY:

All

SENSITIVITY:

Transfected Only

MW (kDa):

Source/Isotype:

Rabbit IgG

Product Information

Product Usage Information

Application	Dilution
Flow Cytometry (Live)	1:50

Storage

Supplied in PBS (pH 7.2), less than 0.1% sodium azide, and 2 mg/mL BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Specificity / Sensitivity

Whitlow/218 Linker F(ab')₂ fragment (Alexa Fluor^® 647 Conjugate) recognizes exogenously expressed levels of scFv-based CARs containing a Whitlow/218 linker. This F(ab')₂ fragment contains no F_c domain and will not exhibit cross-reactivity with F_c receptors.

Species Reactivity:

All Species Expected

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues of the Whitlow/218 linker peptide.

Product Description

This Cell Signaling Technology^® F(ab’)₂ fragment lacks an F_c domain, and will not exhibit reactivity with human F_c receptors. This eliminates the need for F_c blocking and reduces false positives in assays containing human monocytes or other cells with high expression of F_c receptors.

The F(ab')₂ fragment is generated by enzymatic digestion of the full-length IgG clone coupled with chromatography to remove both the F_c fragment and the intact IgG. The F(ab’)₂ fragment is conjugated to Alexa Fluor^® 647 fluorescent dye under optimal conditions.

Background

The Whitlow/218 peptide is a commonly used linker that connects the variable heavy (VH) domain and variable light (VL) domain of single-chain variable fragments (scFvs) and chimeric antigen receptors (CARs) that utilize an extracellular domain scFv for target antigen recognition. The Whitlow/218 linker affords the scFv with enhanced resistance to proteolysis and aggregation (1).

Human F_c receptors exhibit strong cross-reactivity with rabbit IgG, and are expressed at high levels on cells of myeloid lineage (e.g., monocytes and myeloid-derived cell lines). Fluorescent anti-linker F(ab')₂ fragment conjugates are ideal for use in flow cytometry assays on cells containing F_c receptors, and eliminate the need for an F_c blocking step prior to antibody incubation.

Cell Signaling Technology employs strict quality control procedures to ensure that each conjugate provides optimal specificity and fluorescence.

Whitlow, M. et al. (1993) Protein Eng 6, 989-95.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

FC-L: Flow Cytometry (Live)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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