4 Control RNA HAV RNA, in vitro transcript. 04 655 877 001 50 μl, 200 copies/μl
5 Control Primer Mix,
HAV RNA
HAV RNA upstream
and downstream primer to
amplify a 246 bp fragment.
04 655 877
utilize horseradish peroxidase conjugates.1-4 This
substrate produces a soluble end product that is blue in
color and can be read spectrophotometrically at 370 or
655 nm. The reaction may be stopped with
peroxidase (HRP)
conjugates.1-4 This TMB-HRP reaction produces a
soluble end-product that is blue in color and can be
read spectrophotometrically at 370 nm or 655 nm.
The reaction may be stopped
enzyme assay,
which results in a colorimetric (655 nm) product
proportional to the SMase activity present.
Components
The kit is sufficient for 200 assays in 96 well plates.
Enzyme Mix 2 ea
Catalog
1990) 136:641-655.
Important Note: During shipment, small volumes of product will occasionally become entrapped in the seal of the product vial. For
products with volumes of 200 μL or less
produces a soluble end product that is blue in
color and can be read spectrophotometrically at 370 or
655 nm. The reaction maybe stopped with 2 M H2SO4,
resulting in a yellow solution that is read at 450
a soluble end
product that is blue in color and can be read
spectrophotometrically at 370 or 655 nm. The reaction
may be stopped with 2 M H2SO4, resulting in a yellow
solution that is read at
Schaafsma, A.
Huijsmans, H. Robben, G. Schaart and P. Vooijs. American Journal of Pathology
136:641-655, 1990
2. Immunohistochemical demonstration of Keratin 7 in routinely fixed paraffin-embedded
centrifuging at 200 x g for 10 minutes in each wash.
3 . Incubate the cells in 100 μlof a I:20 dilution of fluorescein-conjugated goat anti-mouse IgG.
4. Wash the cells twice with PBS, centrifuging at 200 x g for
10 minutes.
4. Wash the wells three times, 300 µl per well, with
PBS containing 0.05% TWEEN 20 to remove the
nonbiotinylated, complimentary strand of the PCR
product.
5. Add 200 µl per well of
10 minutes.
4. Wash the wells three times, 300 µl per well, with
PBS containing 0.05% TWEEN 20 to remove the
nonbiotinylated, complimentary strand of the PCR
product.
5. Add 200 µl per well of
10 minutes.
4. Wash the wells three times, 300 µl per well, with
PBS containing 0.05% TWEEN 20 to remove the
nonbiotinylated, complimentary strand of the PCR
product.
5. Add 200 µl per well of
for 5–10 minutes.
4. Wash the wells three times, 300 µl per well,
with PBS + 0.05% Tween-20 to remove the
nonbiotinylated, complimentary strand of the
PCR product.
5. Add 200 µl per well of a hapten
soluble end
product,2 which is blue in color and can be read
spectrophotometrically at 370 nm or 655 nm. The
reaction can be stopped with 2 M sulfuric acid (H2SO4)
and read at 450 nm.
Each
x 5 min
4. Incubate with secondary 1 hr 30 min
antibody
5. Wash the membrane 3 x 10 min 3 x 5 min
6. Add substrate 10 min 10 min
Total time 4 hr 10 min