Safety Methods Database Version 0 9 pot
U. S. ECONOMY 2 0 0 9 pot
... first half
of the 1 890 s, forcing some
16 ,00 0 businesses to fail in 1 893
alone. The following year, as
many as 7 50, 000 workers were on
strike, and the unemployment
rate reached 20 percent.
Farmers ... is
responsible for 20 percent of total
economic output.
The U.S. gross domestic prod-
uct per person was nearly $45 ,00 0
in 200 7, compared to a worldwide
average of $11 ,00 0. The eco...
Glycoprotein methods protocols - biotechnology 048-9-001-013.pdf
... manufacturer’s instructions.
9. Centrifuge the samples at 36 ,00 0 rpm ( 50. 2Ti rotor, starting density 1. 50 g/mL) or 40, 000
rpm ( 70. 1Ti rotor, starting density 1.52 g/mL) at 15°C for 72 96 h (see Note 7). ... Samples are centrifuged at 36 ,00 0
rpm ( 50. 2Ti rotor) or 40, 000 rpm ( 70. 1Ti rotor) at 15°C for 72 96 h (see Note 7). These
conditions give gradients of approx 1.25–1. 60...
Glycoprotein methods protocols - biotechnology 048-9-027-044.pdf
... with 0. 000 1% H
2
O
2
in TBS,
pH 7.6.
Histological Methods for Detection of Mucin 35
10. Antigen retrieval solutions: 0. 001 0. 01 M citric acid, pH 6 .0 (pH 2.5–6 .0) , 0. 5 M Tris-
HCl, pH 9. 5– 10. 0 ± ... detergent such as Tween- 20 (0. 05 0. 5%) or Triton X- 100 (0. 1–1%) to the
wash buffers.
c. Increasing the osmolarity of the buffers used for antibody dilution and washing.
Hi...
Glycoprotein methods protocols - biotechnology 048-9-045-055.pdf
... tetramethylbenzidine (TMB) (0. 01
mg/mL, A
450nm
) in Na acetate with 0. 01% H
2
O
2
(pH 6 .0) ; p-nitrophenyl phosphate (PNPP)
(1 mg/mL, A
405 nm
) in 10 mM diethanolamine with 0. 5 mM MgCl
2
(pH 9. 5).
3. Methods
3.1. ... buffer, pH 9. 6; phosphate-buffered saline
(PBS) = 0. 05 M phosphate, 0. 9% (w/v) NaCl, 0. 02% KCl, pH 7.2; Tris-buffered saline
(TBS) = 0. 01 M Tris-HCl, 0...
Glycoprotein methods protocols - biotechnology 048-9-057-064.pdf
... mucous layer. A continuous mucous coat of mean thickness
200 – 300 µm is seen in rat stomach and duodenum (21,22), and a mean thickness of
800 µm is seen in the colon (23). Strong suction will remove ... prepared by dissolving 10 g of gelatin in 60 mL of
distilled water mixed with 2 50 mg of phenol in 70 mL of glycerol.
8. Poly-
L
-lysine-coated slides: slides are immersed in 0. 01% (w...
Glycoprotein methods protocols - biotechnology 048-9-065-073.pdf
... 21).
20. Homogenization buffer (pH 7.5, 0 C): 50 mM Tris-HCl, 5 mM EDTA, 1% Triton X- 100
(BDH, Poole, UK), 10 mM iodacetamide, 100 µg/mL soybean trypsin inhibitor, 10 µg/mL
pepstatin A, aprotinin 1% ... filter paper.
19. Molecular weight marker: nonreduced monomeric and dimeric rat gastric mucin precur-
sors, molecular mass 300 and 600 kDa, respectively (8) (see Chapters 20 and 21)....
Glycoprotein methods protocols - biotechnology 048-9-075-085.pdf
... Electrophoresis buffer: 40 mM Tris-acetate, 1 mM EDTA, pH 8 .0, containing 0. 1% (w/v)
sodium dodecyl sulfate.
4. Loading buffer: electrophoresis buffer containing 30% (v/v) glycerol and 0. 002 % (w/v)
bromophenol ... a flow rate of 2 40 µL/min with
0. 1% (v/v) TFA (5 min) followed by a linear gradient of 0 50% (v/v) acetonitrile in 0. 1%
(v/v) TFA ( 30 min) using the Pharmacia SMART...
Glycoprotein methods protocols - biotechnology 048-9-087-096.pdf
... (M) ρ (g/mL) η
rel
6 .00 1.145 1.6 20
6.25 1.1 50 1.685
6. 50 1.156 1.7 70
6.75 1.162 1.845
7 .00 1.168 1 .92 5
7.25 1.174 2 .03 0
7. 50 1.1 80 2.125
7.75 1.186 2.2 30
8 .00 1. 192 2. 400
a
The values for ρ ... gradient (12 mL) and centrifuged at 202 ,00 0 g
average ( 40, 000 rpm) for 2.5 h at 15°C in a Beckman SW 40 Ti swing-out rotor. The gradient
was emptied from the top into 0....
Glycoprotein methods protocols - biotechnology 048-9-111-119.pdf
... 203 ,00 2,4 80 1 69, 595 , 90 4 2 09 5.1
Met 144 , 09 6,656 29, 927,3 80 52 1.3
Ile 202 ,357,184 132,131,184 163 4 .0
Leu 201 ,99 3 ,02 4 2 70, 782,624 335 8.2
Phe 206 ,174,464 104 ,213 ,9 60 126 3.1
Lys 292 ,284,352 1 09 ,133,576 ... pmol
b
Composition
c
Asx 258, 795 ,7 60 404 ,2 79, 488 391 9. 6
Glx 215,7 20, 688 532,183,264 617 15.1
Hydroxy-Pro 2 49, 767,456 243,187, 696 243 0. 0
S...
Glycoprotein methods protocols - biotechnology 048-9-121-128.pdf
... at 80 C and 50 C.
2. Speed-Vac™ vacuum centrifuge (Savant Instruments, NY)
3. Beckman LF3 600 protein sequenator, or Hewlett Packard G 100 0A protein sequencer, or
PE Biosystem’s Procise
™
.
3. Methods
3.1. ... 1 194 –1 201 .
6. Laursen, R. A. ( 197 1) Solid-phase Edman degradation: an automated peptide sequencer.
Eur. J. Biochem. 20, 89 102 .
7. Laursen, R. A., Lee, T. T., Dixon, J. D., an...