PerkinElmer
check quantity

TNFα (human) AlphaLISA Biotin-Free Detection Kit, 5,000 Assay Points

The AlphaLISA Human TNFα Biotin-Free Detection Kit is designed for the quantitative detection of human TNFα in serum, cell culture medium, and other samples types using a homogeneous (no wash steps, no separation steps) assay. The biotin-free kit uses anti-DIG (anti-Digoxin) Donor beads instead of streptavidin Donor beads, which makes the kit compatible with high-biotin culture media and other sample types that contain high levels biotin (including brain/liver tissue extracts, milk and eggs).

For research use only; not for diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).

Part Number
Unit Size
List Price
Your Price
Quantity
AL325HV
100 assay points
828.00 USD
 
more
AL325C
500 assay points
1811.00 USD
 
more
AL325F
5,000 assay points
12000.00 USD
 
more
Buy Now

Please enter valid quantity

Please log in to add favorites.

NULL OR EMPTY CART

Overview

Formats:

  • Our HV (100 assay point) kits allow you to run 100 wells in 96-well format, using a 100 µL reaction volume (10 µL of sample).
  • Our 500 assay point kit allows you to run 500 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
  • Our 5,000 assay point kit allows you to run 5,000 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).

Features and Benefits:

  • No-wash steps, no separation steps
  • Compatible with high biotin samples, including RPMI and high-biotin tissue extracts
  • ELISA alternative technology
  • Sensitive detection
  • Broad sample compatibility
  • Results in less than 3 hours

Tumor Necrosis Factor Alpha is a multifunctional proinflammatory cytokine synthesized mainly by nucleated blood cells as a 233 aa type II transmembrane protein which is cleaved by ADAM17 between aa76-77 to form a soluble homotrimeric complex. TNFα plays a role in lipid metabolism, coagulation and endothelial function and has been associated with cancer, infection and inflammation (including inflammatory bowel disease), ischemia/reperfusion injury and heart failure, and insulin resistance. This kit is used when the analyte is in a matrix containing high levels of biotin, such as can be found in cell culture media such as RPMI, or milk.

AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA biotin-free assay, a DIG-labeled anti-analyte antibody binds to the anti-DIG-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.

Specifications

Assay Target TNFα
Assay Target Class Cytokine
Automation Compatible Yes
Detection Method Alpha
Experimental Type In vitro
Product Brand Name AlphaLISA
Shipping Condition Blue Ice
Therapeutic Area Inflammation
Unit Size 5,000 assay points
Resources, Events & More
  • All

Application Note

Rapid, No-wash Measurement of Immune Checkpoint Molecules and Cytokines in Co-cultures of Immune Cells and Cancer Cell Lines

Breast cancer tumors can adapt to immune cell infiltration by responding to the increased concentration of interferon gamma (IFN-ɣ) and other cytokines secreted by subsets of T lymphocytes with the upregulation of the immune checkpoint proteins such as Programmed cell death ligand 1 (PD-L1). These checkpoint proteins allow the tumors to evade immune targeting and reduce the immune response, thus promoting tumor progression.

In this application note, you will learn:

  • How exogenous addition of IFN-ɣ effects the expression of PD-L1 and secretion of several cytokines in cultures of HCC38 cells (a triple-negative breast cancer cell line)
  • How co-culturing activated immune cells and breast cancer cells stimulates differential expression of some immune checkpoint and inflammatory biomarkers compared to culturing cells alone with PBMC-conditioned media
  • How to rapidly measure multiple biomarkers in cell culture supernatant and lysates from the same wells of a culture dish to examine protein expression profiles from cancer and immune cell culture models using AlphaLISA® assays together with ATPlite 1step and the EnVision® multimode plate reader.

PDF 2 MB
Using AlphaLISA Biomarker Kits to Assess Effects of PBMC-Conditioned Media on 3D and 2D Cell Culture Models of Breast Cancer

Various cytokines are secreted during an active immune response that can have modulatory effects on target cell populations, including interferon gamma (IFN-ɣ), tumor necrosis factor alpha (TNFa) and several interleukins.

In this application note, you will learn how we investigated:

  • The effects of secreted factors on a triple-negative breast cancer cell line by treatment with conditioned media from activated PBMCs
  • Whether the modulatory effects of cytokines secreted by infiltrating immune is different when studied in 3D cell cultures
  • How to detect and quantify multiple immune-modulated proteins from the same wells in complex cell models grown in 3D spheroid microplates and traditional 2D cell culture using AlphaLISA® technology and EnVision® multimode plate reader in a workflow with ATPlite 1step and Opera® Phenix high-content screening system

PDF 2 MB

Brochure