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PathScan® RP G4S Linker/CD3ζ CAR Sandwich ELISA Kit #20418

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  • ELISA
ELISA Image 1: PathScan® RP G4S Linker/CD3ζ CAR Sandwich ELISA Kit
Figure 1. The PathScan® RP G4S Linker/CD3ζ CAR Sandwich ELISA Kit specifically detects an scFv-based CAR containing a G4S linker. Cell extracts were made from Jurkat cells engineered to stably express an scFv-based Anti-CD20 CAR containing a G4S linker (Jurkat G4S), and Jurkat cells engineered to stably express an scFv-based Anti-CD20 CAR containing a Whitlow/218 linker (Jurkat Whitlow). The relationship between lysate protein concentration from Jurkat, Jurkat G4S, and Jurkat Whitlow cells and the absorbance at 450 nm using the PathScan® RP G4S Linker/CD3ζ CAR Sandwich ELISA Kit #20418 is shown.
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To Purchase # 20418**

Important Ordering Details

Custom Ordering Details:

If kit quantities from the same lot are needed in unlisted sizes, contact us for processing time and pricing.

Looking for this ELISA kit in a 384-well format? Inquire for availability, processing time, and pricing.

Supporting Data

REACTIVITY All
Application Key:
  • ELISA-ELISA 
Species Cross-Reactivity Key:
  • All-All Species Expected 
  • Product Includes
  • Related Products
Product IncludesVolumeSolution Color
CD3ζ Rabbit mAb Coated Microwells #4056096 tests
G4S Linker Rabbit Detection mAb #557141 eaRed (Lyophilized)
HRP Diluent #135155.5 mlRed
TMB Substrate #700411 ml
STOP Solution #700211 ml
Sealing Tape #545032 ea
ELISA Wash Buffer (20X) #980125 ml
Cell Lysis Buffer (10X) #980315 ml

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.

The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Product Information

Product 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.

Protocol

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.

Species Reactivity:

All Species Expected

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 or 20-mer (G4S)4 within 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).

限制使用

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