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pHSense Human IgG4 Isotype Control, 50 µg
pHSense Human IgG4 Isotype Control, 50 µg
generic vial control green cap
pHSense™ Human IgG4 Negative Control is a purified recombinant antibody designed to serve as an isotype control when working with fully human, humanized, or chimeric antibodies in combination with pHSense reagents.
| Feature | Specification |
|---|---|
| Application | 内部化 |
pHSense™ Human IgG4 Negative Control is a purified recombinant antibody designed to serve as an isotype control when working with fully human, humanized, or chimeric antibodies in combination with pHSense reagents.
Product variant
Unit Size: 1 vial
Part #:
81HUIGG4NC
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).
pHSense Human IgG4 Isotype Control, 50 µg
generic vial control green cap
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Product information
Overview
pHSense Eu Fab reagents are designed to monitor antibody or anti-drug conjugate (ADC), as well as receptor- and membrane protein-mediated internalization in real time. To confidently attribute observed effects to specific antigen binding, it is crucial to include proper controls. Test antibodies may engage in non-specific interactions - particularly when their Fc regions bind to Fc receptors on cells - which can contribute to background signal. This is where negative control antibodies become essential in pHSense assays. They help distinguish true signal from background noise by revealing non-specific binding effects, thereby enhancing the reliability of your internalization measurements.
The pHSense Human IgG4 Negative Control is a full-length recombinant IgG4 antibody, composed of both heavy and light chains. It serves as an ideal isotype control when evaluating fully human, humanized, or chimeric antibodies in conjunction with pHSense reagents.
Assay validation
Human IgG4 negative control application in pHSense-based internalization monitoring assay
Tag-Lite® GLP-1R cells (stable cell line, Part #: C1SU1GLP1, Revvity) were seeded in a 96-well white culture-treated plate at a density of 80,000 cells per well in complete culture medium, and incubated overnight at 37°C with 5% CO₂. Dulaglutide, a GLP-1R agonist composed of a GLP-1 analog covalently linked to the Fc fragment of human IgG4, and the Human IgG4 Negative Control were pre-incubated at room temperature for 30 minutes with pHSense Eu Fab Anti-Human IgG (Part #: 81HUEU1AA, Revvity) at a 1:2 molar ratio in cell culture medium.
The Fab–Dulaglutide and Fab–IgG4 Negative Control mixtures were then serially diluted in cell culture medium, and 50 µL of each dilution were added to the cells, followed by a 20-minute incubation at 37°C with 5% CO₂.
The fluorescence signal was recorded using a TRF-compatible plate reader. As expected, Dulaglutide induced a dose-dependent increase in signal, while no signal increase was observed with the Human IgG4 Negative Control.
Specifications
| Application |
Internalization
|
|---|---|
| Brand |
pHSense
|
| Detection Modality |
pH sensitive dye
|
| Product Group |
Fluorescent Reagent
|
| Shipping Conditions |
Shipped in Dry Ice
|
| Target Class |
ADCs
Antibodies
Cell surface proteins
GPCRs
|
| Target Species |
Human
|
| Technology |
TRF
|
| Unit Size |
1 vial
|
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Brochure
pHSense reagents product list
Discover pHSense™ probes – plate reader-compatible reagents with time-resolved fluorescence (TRF) readout, specifically developed...
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