Currently
all our antibodies are purified from sheep serum. The
US Department for Agriculture (USDA) demands an Import
Permit for farm animal products, includes purified antibodies.
It requires from you to fill in APHIS
Form 16-3 and submit it to the USDA (USDA
Info sheet). There is a $ 94 fee to USDA attached
to this. It takes 4 to 8 weeks to obtain the import
permit from the USDA. We apologize for the inconvenience,
but there is nothing we can do to take this burden from
you. To our knowledge no other country has this requirement.
Since the beginning of 2008 it seems the USDA is increasingly
unhelpful with regards to this permit.
The
Importer is: you, the customer.
The
Producer/Shipper is:
Kinasource Ltd
James Black Centre
Dow Street
Dundee DD1 5EH
Scotland, UK
The
US port of entry is: "as applicable"
Mode
of Transportation is "by AIR"
The
material imported is described as:
The material is: antibody affinity purified
from sheep serum.
Sheep are housed at Diagnostics Scotland, Barnhouse
Farm, Bonnybridge, Scotland.
Material was processed at Division of Signal Transduction
Therapy, Faculty of Life
Sciences, MSI/WTB complex, University of Dundee, Dundee
DD1 5EH.
The material has been purified by affinity chromatography
against the peptide or protein
immunogen and the samples contain no other animal products
such as bovine serum albumin.
2.TREATMENT
OF MATERIAL PRIO PRIOR TO IMPORTATION INTO THE U.S.
(Processing/purification methods, including time at
specific temperatures, pH,other treatments, disease
safeguard etc.)
Antibodies have been derived only from
animals considered to be free of evidence of disease.
Antibodies do not contain any animal products such as
albumin and are for laboratory use only. The materials
are non-hazardous and non-contagious.
Purification was carried out using the following method.
1.Wash the column with 20 column volumes of 25 mM Tris/HCL
(pH7.5) + 0.5M NaCl, then equilibrate the column with
5 column volumes of 25mM Tris/HCL pH7.5/1% Triton X-100
(loading buffer).
2. Pre-heat the serum for 20 min at 56°C and filter
it through 0.45 mm.
3. Dilute the anti-serum to 100 ml with distilled and
then 1:1 in 50mM Tris/HCL pH7.5 + 2% Triton X-100. Load
this diluted serum onto the equilibrated column 4 times.
Keep the load. Store at 40C
4. Wash the column with at least 20 beads volumes of
25mM Tris/HCl (pH7.5) containing 0.5M NaCl and then
with at least 20 beads volumes of 25mM Tris/HCl pH7.
Check the OD280 nm. It should be <0.04. Keep the
wash buffer at 40C.
5. Elution. Elute bound Abs by passing 10 beads volumes
of 50mM Glycine/HCl pH2.2 through the column. Collect
the eluate in glass tubes containing 1:10 1M Tris/HCl
(pH8.0). Collect 5ml fractions. Check the protein concentration
in each fraction, by measuring the OD at 280 nm (by
using the 1 cm Quartz cuvette, an OD 280 = 1.35 corresponds
to an Ab concentration of 1 mg/ml). Pool all the fractions
with [Ab] > 0.2 mg/ml.
6. Dialyse overnight against PBS, concentrate if necessary
and store at –20°C.
7. Just after elution, neutralise the column with 25
mM Tris/HCL (pH7.5). Store at 4°C in
25 mM Tris/HCL (pH7.5) + 0.15 M NaCl, 0.02% Na Azide
for protein columns or in 20% ethanol for peptide columns.
8. If you have to use the Dephospho column, pre-equilibrate
the column with 20 column
volumes of PBS.
9. Load at least 3 times the column with the pooled,
dialysed and concentrated ‘eluate’
from the phospho column, collect to flow through corresponding
to the Phospho-specific Ab. Measure the Ab concentration
and concentrate if necessary. Aliquot as appropriate,
and store at –20°C.
10. Elute, as for step 5, these fractions are the Dephospho
antibody.
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