Activated Immunobead® Matrix - Irvine Scientific

INTENDED USE
200 mg of Immunobead matrix is supplied lyophilized and contains
0.01% NaN 3 . Re-hydration with 20 mL of distilled water produces a 10
mg/mL suspension in the recommended coupling buffer. Do not freeze
after hydration. Store at 4°C.
Antibody coupling is done at pH 6.3; however, coupling of other types of
proteins is done at a pH close to the isoelectric point of the protein. For
example, human serum albumin (isoelectric point = 4.8) is coupled to
the Immunobead matrix in 0.01 M pyridine buffer, pH 4.8. It is essential
that the coupling be run at low ionic strength (conductivity 1,000 µohms).
DIRECTIONS FOR USE
A. Buffers
1. Coupling buffer (0.003 M phosphate, pH 6.3).
Prepare a 0.3 M stock solution of monobasic potassium phosphate
(KH 2 PO 4 anhydrous, m.w. = 136.1) by dissolving 40.8 g of KH 2 PO 4
in 1 liter H 2 O. Dilute 10mL stock 0.3 M KH 2 PO 4 with 800 mL water.
Adjust pH to 6.3 with 0.1 M KOH. Dilute to 1.0 liter.
2. Phosphate buffered saline (PBS) (0.01 M KH 2 PO 4 , 0.15 M NaCl, pH
7.2 with KOH).
Dissolve 478 g sodium chloride (NaCl) and 0.1 g NaN 3 in 500 mL
water. Add 33.3 mL of a 0.3 M KH 2 PO 4 stock solution. Adjust to pH
7.2 with 1 M KOH. Dilute to 1.0 liter with water
3. 1.4 M sodium chloride (NaCl) – PBS.
Dissolve 73.22 g NaCl in 700 mL PBS. Dilute to 1.0 liter with PBS
and adjust pH to 7.2.
If high background becomes a problem, the following buffer is
suggested in place of 1.4 M NaCl – PBS: 5M guanidine hydrochloride
(GuHCl) – PBS.
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