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Emina Vorkapic, Maggie Folkesson, Kerstin Magnell, Mohammad Bohlooly-Y, Toste Länne, Dick Wågsäter
ADAMTS-1 in abdominal aortic aneurysm
www.plosone.org
www.plosone.org
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/F7 1 Tf
-0.004 Tc
(80%)Tj
2.7042 0 Td
(power.)Tj
2.948 0 Td
(Animals)Tj
3.5773 0 Td
(were)Tj
2.126 0 Td
(housed)Tj
3.0954 0 Td
(in)Tj
1.0318 0 Td
(groups)Tj
2.9707 0 Td
(in)Tj
1.0375 0 Td
(pathogen)Tj
3.94 0 Td
(free)Tj
1.7291 0 Td
(\(SPF\))Tj
2.4605 0 Td
(facility)Tj
2.897 0 Td
(in)Tj
1.0374 0 Td
(IVC)Tj
-35.7672 -1.2983 Td
(enriched)Tj
3.7304 0 Td
(cages)Tj
2.33 0 Td
(and)Tj
1.7292 0 Td
(maintained)Tj
4.7905 0 Td
(on)Tj
1.2756 0 Td
(a)Tj
0.6519 0 Td
(12:12)Tj
2.3471 0 Td
(hour)Tj
2.1543 0 Td
(light/dark)Tj
4.1839 0 Td
(cycle.)Tj
-21.9967 -1.3039 Td
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1.2076 0 Td
(eight)Tj
2.1996 0 Td
(weeks)Tj
2.6079 0 Td
(of)Tj
1.0148 0 Td
(age,)Tj
1.7688 0 Td
(AAA)Tj
2.2734 0 Td
(was)Tj
1.6951 0 Td
(induced,)Tj
3.6963 0 Td
(in)Tj
1.0318 0 Td
(randomly)Tj
4.1329 0 Td
(chosen)Tj
3.0104 0 Td
(mice,)Tj
2.3753 0 Td
(by)Tj
1.1849 0 Td
(chronic)Tj
3.2882 0 Td
(infusion)Tj
3.5319 0 Td
(of)Tj
-36.2151 -1.2982 Td
(1000)Tj
2.1203 0 Td
(ng/kg/min)Tj
4.4901 0 Td
(AngII)Tj
2.5965 0 Td
(\(Cat.no.9525,)Tj
5.5899 0 Td
(Sigma)Tj
2.6815 0 Td
(Aldrich,)Tj
3.498 0 Td
(St.)Tj
1.2188 0 Td
(Louis,)Tj
2.6419 0 Td
(USA\))Tj
2.4605 0 Td
(via)Tj
1.3718 0 Td
(mini-osmotic)Tj
5.6579 0 Td
(pumps)Tj
-34.3272 -1.2983 Td
(\(Model)Tj
3.1521 0 Td
(1004,)Tj
2.3414 0 Td
(Alzet,)Tj
2.5058 0 Td
(CA,)Tj
1.7858 0 Td
(USA\))Tj
2.4492 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0421 0 Td
(previously)Tj
4.3484 0 Td
([)Tj
0.83 0.64 0.02 0 k
(11)Tj
0 g
(].)Tj
2.0692 0 Td
(Pumps)Tj
2.9991 0 Td
(were)Tj
2.1202 0 Td
(placed)Tj
2.7836 0 Td
(under)Tj
2.6135 0 Td
(anesthe-)Tj
-34.2252 -1.3039 Td
(sia)Tj
1.2813 0 Td
(using)Tj
2.3867 0 Td
(isoflurane.)Tj
4.422 0 Td
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5.9924 0 Td
(pain)Tj
1.9843 0 Td
(was)Tj
1.7008 0 Td
(treated)Tj
2.999 0 Td
(with)Tj
1.9956 0 Td
(buprenorphine)Tj
6.2757 0 Td
(at)Tj
0.9525 0 Td
(a)Tj
0.6576 0 Td
(dose)Tj
2.0353 0 Td
(of)Tj
1.0148 0 Td
(0.1)Tj
1.4003 0 Td
(mg/)Tj
-35.0983 -1.2983 Td
(kg)Tj
1.1792 0 Td
(body)Tj
2.2167 0 Td
(weight,)Tj
3.1124 0 Td
(injected)Tj
3.3902 0 Td
(subcutaneously)Tj
6.3666 0 Td
(twice)Tj
2.3187 0 Td
(per)Tj
1.5307 0 Td
(day)Tj
1.6328 0 Td
(for)Tj
1.3889 0 Td
(3)Tj
0.6974 0 Td
(days.)Tj
2.2223 0 Td
(A)Tj
0.9014 0 Td
(group)Tj
2.6078 0 Td
(of)Tj
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1.0148 0 Td
[()12()4()]TJ
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 525.6 499.3511 cm
BT
/F14 1 Tf
1 TL
-0.0053 Tc
0 0 Td
[()]TJ
ET
Q
q
1 j
1 J
0 w
10 0 0 10 535.011 494.9291 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(mice)Tj
2.1543 0 Td
(were)Tj
-35.6539 -1.2982 Td
(infused)Tj
3.1635 0 Td
(with)Tj
1.9955 0 Td
(0.9%)Tj
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2.3017 0 Td
(and)Tj
1.7291 0 Td
(were)Tj
2.1203 0 Td
(used)Tj
2.0636 0 Td
(as)Tj
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(control)Tj
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(mice.)Tj
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3.8381 0 Td
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(diet)Tj
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(and)Tj
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(water)Tj
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(allowed)Tj
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3.2882 0 Td
[()6(\r)]TJ
1.2019 0 Td
[()12()14()11()4(\r)12()12()]TJ
/F7 1 Tf
3.2655 0 Td
(throughout)Tj
4.7791 0 Td
(the)Tj
1.4684 0 Td
(whole)Tj
2.6135 0 Td
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(and)Tj
1.7291 0 Td
(mice)Tj
2.1543 0 Td
(were)Tj
2.126 0 Td
(monitored)Tj
4.49 0 Td
(daily)Tj
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1.3833 0 Td
(signs)Tj
2.2222 0 Td
(of)Tj
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(discomfort.)Tj
4.8132 0 Td
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2.2847 0 Td
(28)Tj
1.1736 0 Td
(days,)Tj
2.2223 0 Td
(mice)Tj
2.1543 0 Td
(were)Tj
2.1203 0 Td
(sacrificed.)Tj
4.235 0 Td
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1.7801 0 Td
(aorta)Tj
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(removed)Tj
3.7474 0 Td
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1.7291 0 Td
(fixated)Tj
2.9254 0 Td
(in)Tj
1.0373 0 Td
(RNAla-)Tj
-34.1855 -1.2983 Td
(ter)Tj
1.3096 0 Td
(for)Tj
1.389 0 Td
(24)Tj
1.1735 0 Td
(hours)Tj
2.5115 0 Td
(thereafter)Tj
4.0819 0 Td
(frozen)Tj
2.7722 0 Td
(in)Tj
1.0318 0 Td
(-70C)Tj
2.5795 0 Td
(for)Tj
1.3833 0 Td
(gene)Tj
2.0807 0 Td
(expression)Tj
4.4673 0 Td
(analysis)Tj
3.3279 0 Td
(or)Tj
1.0942 0 Td
(snap)Tj
2.0806 0 Td
(frozen)Tj
2.7721 0 Td
(for)Tj
1.389 0 Td
(pro-)Tj
-35.4441 -1.2982 Td
(tein)Tj
1.7575 0 Td
(analysis.)Tj
3.5489 0 Td
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1.8822 0 Td
(of)Tj
1.0148 0 Td
(aorta)Tj
2.2677 0 Td
(was)Tj
1.6951 0 Td
(fixed)Tj
2.194 0 Td
(in)Tj
1.0375 0 Td
(formaldehyde)Tj
5.7713 0 Td
(and)Tj
1.7291 0 Td
(embedded)Tj
4.3994 0 Td
(in)Tj
1.0375 0 Td
(paraffin)Tj
3.3675 0 Td
(for)Tj
1.3833 0 Td
(histology)Tj
-33.0858 -1.3039 Td
(and)Tj
1.7291 0 Td
(analysis)Tj
3.3279 0 Td
(of)Tj
1.0148 0 Td
(aneurysm)Tj
4.1612 0 Td
(formation.)Tj
4.4844 0 Td
(The)Tj
1.7802 0 Td
(study)Tj
2.3924 0 Td
(was)Tj
1.6951 0 Td
(approved)Tj
3.9741 0 Td
(by)Tj
1.1849 0 Td
(the)Tj
1.4683 0 Td
(local)Tj
2.075 0 Td
(ethical)Tj
2.8403 0 Td
(committee)Tj
4.5013 0 Td
(in)Tj
-36.629 -1.2983 Td
(Linko)Tj
0 Tc
1.9106 0 Td
()Tj
-0.004 Tc
0.4478 0 Td
(ping,)Tj
2.2394 0 Td
(Sweden)Tj
3.3052 0 Td
(\(4713\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 377.4614 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Immunohistochemi)Tj
8.3008 0 Td
(stry)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 360.4535 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
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7.9369 0 Td
(human)Tj
3.0784 0 Td
(abdominal)Tj
4.5127 0 Td
(aortas)Tj
2.6249 0 Td
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2.2053 0 Td
(four)Tj
1.9163 0 Td
(patients)Tj
3.3675 0 Td
(were)Tj
2.1203 0 Td
(sectioned)Tj
4.0025 0 Td
(\(5)Tj
/F15 1 Tf
1.0375 0 Td
[()]TJ
/F7 1 Tf
(m\))Tj
1.8822 0 Td
(and)Tj
-34.6845 -1.2982 Td
(rehydrated)Tj
4.5807 0 Td
(in)Tj
1.0318 0 Td
(several)Tj
2.9367 0 Td
(changes)Tj
3.4015 0 Td
(of)Tj
1.0148 0 Td
(ethanol)Tj
3.2088 0 Td
(and)Tj
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ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 429.4488 351.8929 cm
BT
/F16 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 439.0866 347.4708 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
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(\(Sakura)Tj
3.2995 0 Td
(Finetek,)Tj
3.4299 0 Td
(Leiden,)Tj
3.1747 0 Td
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-33.8113 -1.2982 Td
(Netherlands\).)Tj
5.6806 0 Td
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5.21 0 Td
(peroxidase)Tj
4.5128 0 Td
(activity)Tj
3.1181 0 Td
(was)Tj
1.7007 0 Td
(quenched)Tj
4.1272 0 Td
(by)Tj
1.1848 0 Td
(treatment)Tj
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(with)Tj
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(3%)Tj
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(peroxide)Tj
3.719 0 Td
(for)Tj
1.389 0 Td
(5)Tj
0.6973 0 Td
(min)Tj
1.8539 0 Td
(followed)Tj
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(by)Tj
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(incubation)Tj
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(in)Tj
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(5%)Tj
1.4513 0 Td
(blocking)Tj
3.685 0 Td
(rabbit)Tj
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(or)Tj
1.0999 0 Td
(goat)Tj
1.9218 0 Td
(serum)Tj
2.7269 0 Td
(albumin)Tj
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(solu-)Tj
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2.0636 0 Td
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(were)Tj
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(then)Tj
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(incubated)Tj
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(human)Tj
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(primary)Tj
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(antibodies)Tj
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(against)Tj
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(ADAMTS-1)Tj
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(\(2)Tj
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1.0375 0 Td
[()]TJ
/F7 1 Tf
(g/)Tj
-35.994 -1.2983 Td
(ml,)Tj
1.508 0 Td
(Cat.)Tj
1.8425 0 Td
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(AF5867,)Tj
3.5603 0 Td
(R&D)Tj
2.2847 0 Td
(Systems,)Tj
3.634 0 Td
(Inc,)Tj
1.7574 0 Td
(Minneapolis,)Tj
5.4595 0 Td
(USA\),)Tj
2.6815 0 Td
(CD68)Tj
2.5738 0 Td
(\(0.1)Tj
/F15 1 Tf
1.7348 0 Td
[()]TJ
/F7 1 Tf
(g/ml,)Tj
2.812 0 Td
(Cat.)Tj
1.8425 0 Td
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(NCL-CD68-KP1,)Tj
7.1433 0 Td
(Leica)Tj
2.3073 0 Td
(Microsystems,)Tj
5.9811 0 Td
(Newcastle,)Tj
4.4787 0 Td
(UK\))Tj
1.9673 0 Td
(and)Tj
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1.7347 0 Td
()Tj
/F7 1 Tf
(-actin)Tj
3.1067 0 Td
(\(0.6)Tj
1.7405 0 Td
(mg/ml,)Tj
3.1237 0 Td
(cat.)Tj
1.6044 0 Td
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-33.1877 -1.2982 Td
(A5228,)Tj
3.0331 0 Td
(clone)Tj
2.364 0 Td
(1A4,)Tj
2.0806 0 Td
(Sigma-Aldrich\))Tj
6.4233 0 Td
(at)Tj
0.9524 0 Td
(4C)Tj
1.7518 0 Td
(overnight)Tj
4.0819 0 Td
(followed)Tj
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(by)Tj
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(secondary)Tj
4.2746 0 Td
(biotinylated)Tj
4.9947 0 Td
(rabbit)Tj
-34.7865 -1.2983 Td
(anti-sheep)Tj
4.371 0 Td
(IgG)Tj
1.7405 0 Td
(or)Tj
1.0998 0 Td
(goat)Tj
1.9219 0 Td
(anti-mouse)Tj
4.7565 0 Td
(IgG)Tj
1.7404 0 Td
(antibody)Tj
3.7644 0 Td
(\(Dako,)Tj
2.9594 0 Td
(Stockholm,)Tj
4.745 0 Td
(Sweden\).)Tj
3.8721 0 Td
(Avidin-biotin)Tj
-30.971 -1.3039 Td
(peroxidase)Tj
4.5184 0 Td
(complexes)Tj
4.405 0 Td
(\(Dako\))Tj
3.0727 0 Td
(were)Tj
2.1203 0 Td
(added)Tj
2.6646 0 Td
(followed)Tj
3.651 0 Td
(by)Tj
1.1792 0 Td
(visualization)Tj
5.244 0 Td
(with)Tj
2.0012 0 Td
(3,3)Tj
ET
Q
q
1 j
1 J
0 w
6.9998 0 0 6.9998 500.3149 247.1244 cm
BT
/F13 1 Tf
1 TL
0 0 Td
(0)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 502.696 243.4393 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(-diaminobenzi-)Tj
-30.2681 -1.2982 Td
(dine)Tj
1.9842 0 Td
(tetrahydrochloride)Tj
7.7272 0 Td
(\(Dako\).)Tj
3.2995 0 Td
(All)Tj
1.4003 0 Td
(sections)Tj
3.4073 0 Td
(were)Tj
2.1259 0 Td
(counterstained)Tj
6.1795 0 Td
(with)Tj
1.9956 0 Td
(Mayer's)Tj
3.3731 0 Td
(hematoxylin)Tj
-31.4926 -1.2983 Td
(\(Histolab)Tj
3.9458 0 Td
(Products,)Tj
4.0195 0 Td
(Go)Tj
0 Tc
0.7824 0 Td
()Tj
-0.004 Tc
0.4422 0 Td
(teborg,)Tj
3.016 0 Td
(Sweden\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 190.9984 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Western)Tj
3.9118 0 Td
(blot)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 173.9905 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
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3.0103 0 Td
(aortas)Tj
2.6249 0 Td
(from)Tj
2.2053 0 Td
(AAA)Tj
2.2734 0 Td
(patients)Tj
3.3675 0 Td
(and)Tj
1.7292 0 Td
(non-aneurysmal)Tj
6.7974 0 Td
(control)Tj
3.1238 0 Td
(donors)Tj
3.0387 0 Td
(were)Tj
2.126 0 Td
(homogenised)Tj
5.5955 0 Td
(in)Tj
-35.892 -1.2982 Td
(ice-cold)Tj
3.3902 0 Td
(RIPA)Tj
2.4207 0 Td
(lysis)Tj
1.9106 0 Td
(buffer)Tj
2.6248 0 Td
(containing)Tj
4.5298 0 Td
(50)Tj
1.1735 0 Td
(mM)Tj
1.9276 0 Td
(Tris-HCl)Tj
3.8607 0 Td
(pH)Tj
1.5024 0 Td
(7.4,)Tj
1.6214 0 Td
(150)Tj
1.6497 0 Td
(mM)Tj
1.9333 0 Td
(NaCl,)Tj
2.5228 0 Td
(1)Tj
0.6973 0 Td
(mM)Tj
1.9332 0 Td
(EDTA,)Tj
-33.698 -1.2983 Td
(0.25%)Tj
2.6305 0 Td
(Na-deoxy-cholate,)Tj
7.5514 0 Td
(1%)Tj
1.4514 0 Td
(Triton-100)Tj
4.6034 0 Td
(and)Tj
1.7291 0 Td
(0.1%)Tj
2.1543 0 Td
(SDS,)Tj
2.1317 0 Td
(100)Tj
/F15 1 Tf
1.6497 0 Td
[()]TJ
/F7 1 Tf
(g/ml)Tj
2.5852 0 Td
(phenylmethylsulphonyl)Tj
-26.4867 -1.3039 Td
(fluoride)Tj
3.3788 0 Td
(\(PMSF\))Tj
3.3562 0 Td
(and)Tj
1.7291 0 Td
(protease)Tj
3.5547 0 Td
(inhibitor)Tj
3.7814 0 Td
(cocktail)Tj
3.3165 0 Td
(\(Roche)Tj
3.0614 0 Td
(Diagnostics)Tj
4.8869 0 Td
(Scandinavia,)Tj
5.261 0 Td
(Stockholm,)Tj
-32.326 -1.2983 Td
(Sweden\).)Tj
3.8664 0 Td
(Protein)Tj
3.1974 0 Td
(quantification)Tj
5.8337 0 Td
(of)Tj
1.0148 0 Td
(tissue)Tj
2.4605 0 Td
(extracts)Tj
3.2938 0 Td
(\(30)Tj
/F15 1 Tf
1.5137 0 Td
[()]TJ
/F7 1 Tf
(g)Tj
1.1905 0 Td
(of)Tj
1.0148 0 Td
(protein\))Tj
3.4923 0 Td
(were)Tj
2.1203 0 Td
(separated)Tj
4.0138 0 Td
(under)Tj
-33.012 -1.2982 Td
(reducing)Tj
3.7757 0 Td
(conditions)Tj
4.473 0 Td
(using)Tj
2.3924 0 Td
(412%)Tj
2.9197 0 Td
(gradient)Tj
3.5546 0 Td
(SDS-PAGE)Tj
4.7849 0 Td
(and)Tj
1.7291 0 Td
(transferred)Tj
4.6431 0 Td
(onto)Tj
2.0806 0 Td
(a)Tj
0.652 0 Td
(polyvinylidine)Tj
-31.0051 -1.3039 Td
(difluoride)Tj
4.1725 0 Td
(membrane)Tj
4.5694 0 Td
(\(Amersham)Tj
5.0003 0 Td
(Life)Tj
1.7348 0 Td
(Science,)Tj
3.4299 0 Td
(Amersham,)Tj
4.8869 0 Td
(UK\).)Tj
2.194 0 Td
(Membranes)Tj
5.0003 0 Td
(were)Tj
2.126 0 Td
(blocked)Tj
3.3505 0 Td
(in)Tj
-36.4646 -1.2983 Td
(5%)Tj
1.4513 0 Td
(non-fat)Tj
3.1974 0 Td
(dry)Tj
1.5704 0 Td
(milk)Tj
2.0523 0 Td
(and)Tj
1.7291 0 Td
(incubation)Tj
4.5468 0 Td
(with)Tj
2.0012 0 Td
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