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 11-Dehydrothromboxane B2 RIA test*

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11-dehydro-Thromboxane B2 radioimmunoassay kit (RK-67M)

Intended use:

Thromboxane B2, the stable hydrolysis product of thromboxane A2, is further metabolised to produce 11-dehydro-Thromboxane B2 (11-dehydro-TXB2). In plasma and urine the concentration of 11-dehydro-TXB2 rather than that of TXB2, is accepted to be the relevant parameter of systemic thromboxane production. Extrarenal thromboxane biosynthesis can be most reliably monitored by measuring urinary 11-dehydro-TXB2 concentration. This is clearly assisted by setting up the present RIA system, which combines a high-specific-activity radioiodinated tracer with a sensitive polyclonal rabbit antiplasma. The assay has been thoroughly optimised for determination of urinary 11-dehydro-TXB2 - this is permitted by the elaboration of such a unique selective solid-phase extraction procedure that abolish the requirement of chromatographic purification step.

Assay characteristics:

Assay type:   RIA
Isotope:   125I
Separation:   Magnetic solid phase
Sample type:   Human urine
Sample preparation:   Solid-phase extraction without chromatography
Sample volume:   1 ml
Sample capacity:   40 (duplicate) or 24 (triplicate) unknowns
Time:   One working day
Hands-on time:   3 hours

Special advantages:

- Good sensitivity fitting to urinary concentrations
- Robust and precise assay protocol
- Reasonably short hands-on time
- Reconstituted reagents can be stored for re-use
- Detailed up-to-date theoretical background of thromboxane metabolism
- Homogeneous immunoreactivity obtained with unique solid-phase extraction procedure

Availability: On request

Pack contents:

- 125I-methyltyrosyl-11-dehydro-thromboxane B2 Tracer, in organic solution
- 11-dehydro-thromboxane B2 Standard, 10 ng/ml, lyophilised
- 11-dehydro-thromboxane B2 rabbit Antiserum, lyophilised
- Assay Buffer concentrate
- Magnetizable latex, ready to use
- Full instructions booklet

Assay parameters:

Measuring range:   1.23 - 300 pg/tube  
Typical IC50-value: ~ 25 pg/tube at 50% B / B0  
Sensitivity: ~ 2 pg/tube  
Specificity: See cross reactions below:  
  Thromboxane B2 0.006
  6-keto-PGF1a < 0.010
  Prostaglandin E2 < 0.001
  Prostaglandin F2a < 0.001
  Prostaglandin D2 0.500
  Prostaglandin A2 < 0.001
  Prostaglandin B2 < 0.001
  Prostaglandin F1a < 0.001
  13,14-dihydro-15-keto-PGE2 < 0.001
  13,14-dihydro-15-keto-PGF2a 0.001
  13,14-dihydro-15-keto-PGA2 < 0.010
  13,14-dihydro-6,15-diketo-PGF1a < 0.001
  6-keto-PGE1 < 0.001
  11-dehydro-TXB2 100.00
  2,3-dinor-TXB2 0.040
  2,3-dinor-6-keto-PGF1a < 0.010
  o-hydroxy-hippuric acid < 0.001

11-Dehydrothromboxane B2 RIA test*

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