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Catalog # HK328

TCC ELISA, human, kit

The human terminal complement complex (TCC) ELISA kit is to be used for the in vitro quantitative determination of human TCC in plasma, urine, cerebrospinal fluid, joint fluid and cell culture supernatant samples.

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756.001,223.00

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Harnessing the power of the complement system, the Terminal Complement Complex (TCC),
also known as the Membrane Attack Complex (MAC), stands as a crucial player in our body’s
defense mechanism. When the classical, lectin, and alternative pathways unite, they give rise
to the C5 convertase, which sets the stage for the TCC’s formation. Unlike earlier stages that
depend on enzymatic action, the TCC forms through a cascade of conformational shifts,
beginning with C6 and culminating in the pivotal C7 binding, which fundamentally changes the
complex’s structure.
The TCC is characterized by unique neoantigens, distinct markers that emerge in both the
cellular MAC and the fluid-phase SC5b-9 complex. These markers are absent in the individual
native components, marking the TCC’s presence as a sign of active complement engagement.
Typically, TCC levels in human plasma are negligible, yet an increase signals an immune
response, offering a window into the system’s response to pathogenic threats. By monitoring
TCC levels, we gain invaluable insights into immune system activity, providing a strategic
advantage in both diagnostic and therapeutic realms.

How to analyze complement

For accurate complement analysis, it is important that not only the correct technique is used but also that pre-analytical sample handling is performed in an appropriate manner to avoid erroneous results.

Go to optimal guidelines

Application
The human terminal complement complex (TCC) ELISA kit is to be used for the in vitro quantitative determination of human TCC in plasma, urine, cerebrospinal fluid, joint fluid and cell culture supernatant samples.

Principle
The human TCC ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours. The efficient format of 2 plates with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are captured by a solid bound specific antibody. Biotinylated tracer antibody will bind to captured TCC. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the TCC standards (log). The TCC concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.

Linearity
The linearity of the assay was determined by serially diluting a sample containing 250 mAU/ml human TCC. The diluted samples were measured in the assay. The line obtained a slope of 0.967 and a correlation coefficient of 0.999.

Recovery
Normal human blood samples (plasma), containing baseline levels of human TCC, were spiked with human TCC in concentrations of 2500 and 250 mAU/ml. Samples with and without TCC were incubated for 1 hour at 4° C. Samples were measured by ELISA. Values for TCC ranged between 153% and 128% (mean 140%).

Product type
Assays
Amount
1 x 96 det., 2 x 96 det.
Standard range
8.2 to 2000 mAU/ml
Detection level
8.2 mAU/ml
Working volume
100 µl/well
Species
Human
Cross reactivity
Cynomolgus monkey – Yes, Horse – No, Mouse – No, Pig – Yes, Rabbit – Yes, Rat – No
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Disease
Infectious diseases, Nephrology, Neurological disorders
Safety Data Sheet Assay Manual
HK328
596 kb

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