PathScan^® RP G4S Linker/CD3 zeta CAR Sandwich ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
All

Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

[email protected]

cellsignal.com

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Product Includes	Product #	Quantity	Color	Storage Temp
CD3ζ Rabbit mAb Coated Microwells	40560	96 tests		+4C
G4S Linker Rabbit Detection mAb	55714	1 ea	Red (Lyophilized)	+4C
HRP Diluent	13515	5.5 ml	Red	+4C
TMB Substrate	7004	11 ml		+4C
STOP Solution	7002	11 ml		+4C
Sealing Tape	54503	2 ea		+4C
ELISA Wash Buffer (20X)	9801	25 ml		+4C
Cell Lysis Buffer (10X)	9803	15 ml		-20C

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

Description

The rapid protocol (RP) PathScan^® RP G4S Linker/CD3ζ CAR Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects scFv-based chimeric antigen receptors (CARs) containing a G4S linker and a CD3ζ signaling domain in a reduced assay time of 1.5 hours. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with CARs containing a G4S linker in a single step by using one antibody that recognizes CD3ζ and another antibody that recognizes the G4S linker. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of scFv-based CARs containing a G4S linker and a CD3ζ signaling domain. Learn more about your ELISA kit options here.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

The PathScan^® RP G4S Linker/CD3ζ CAR Sandwich ELISA Kit detects exogenously expressed levels of scFv-based CARs containing a G4S linker and a CD3ζ signaling domain by using one antibody that recognizes CD3ζ and another antibody that recognizes the G4S linker. The kit sensitivity is shown in Figure 1.

Background

The poly-Glycine-Serine (G4S) linker is a type of flexible, unstructured synthetic peptide linker sequence often leveraged to connect 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 linker itself consists of a core pentapeptide sequence, Gly-Gly-Gly-Gly-Ser, that is repeated and commonly found as either a 15-mer (G4S)₃ or20-mer(G4S)₄within scFv-based CARs and scFv fragments. The linker sequence length plays a role in controlling scFv stability and the noncovalent association between the VH and VL domains (1,2).

Background References

Trademarks and Patents

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

PathScan is a registered 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

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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.

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com