Revision 5
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
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) 1:100 - 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #66159.

Specificity / Sensitivity

Whitlow/218 Linker (E3U7Q) Rabbit mAb recognizes exogenously expressed levels of scFv-based CARs containing a Whitlow/218 linker.

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 antibody is tested in-house for indirect flow cytometric analysis in human cells and is expected to react with cell surface expressed CARs of varying specificity, which contain a Whitlow/218 linker within the scFv of the extracellular domain.

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).

  1. 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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 5
#57710

Whitlow/218 Linker (E3U7Q) Rabbit mAb

Flow Cytometry Image 1: Whitlow/218 Linker (E3U7Q) Rabbit mAb Expand Image
Flow cytometric analysis of live pan-CD3+ T cells isolated from human PBMCs and engineered to express an scFv-based Anti-CD19 CAR containing a Whitlow/218 linker, using Whitlow/218 Linker (E3U7Q) Rabbit mAb (right) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Tag Blue fluorescent protein (TagBFP) is co-expressed with the CAR. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Data courtesy of Michael Kvorjak, Lohmueller lab (University of Pittsburgh).
Flow Cytometry Image 2: Whitlow/218 Linker (E3U7Q) Rabbit mAb Expand Image
Flow cytometric analysis of live pan-CD3+ T cells isolated from human PBMCs and engineered to express an scFv-based Anti-CD20 CAR containing a Whitlow/218 linker, using Whitlow/218 Linker (E3U7Q) Rabbit mAb (right) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Tag Blue fluorescent protein (TagBFP) is co-expressed with the CAR. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Data courtesy of Michael Kvorjak, Lohmueller lab (University of Pittsburgh).
Flow Cytometry Image 3: Whitlow/218 Linker (E3U7Q) Rabbit mAb Expand Image
Flow cytometric analysis of live Jurkat cells (blue, negative) or Jurkat cells engineered to stably express an scFv-based Anti-CD19 CAR containing a Whitlow/218 linker (green, positive) using Whitlow/218 Linker (E3U7Q) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Engineered cell line was provided by the Lohmueller Lab, University of Pittsburgh.
Flow Cytometry Image 4: Whitlow/218 Linker (E3U7Q) Rabbit mAb Expand Image
Flow cytometric analysis of live Jurkat cells (blue, negative) or Jurkat cells engineered to stably express an scFv-based Anti-CD20 CAR containing a Whitlow/218 linker (green, positive) using Whitlow/218 Linker (E3U7Q) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Engineered cell line was provided by the Lohmueller Lab, University of Pittsburgh.