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protocol_345.ann
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T1 Action 50 53 Run
E1 Action:T1 Acts-on:T11
T2 Action 73 81 Transfer
E2 Action:T2 Acts-on:T12 Site:T13
T3 Action 100 105 Rinse
E3 Action:T3 Acts-on:T14 Site:T16 Using:T18
T4 Action 179 187 Incubate
E4 Action:T4 Acts-on:T22 Using:T23 Site:T29
T5 Action 282 287 Rinse
E5 Action:T5 Acts-on:T33 Count:T34 Acts-on2:T36
T6 Action 331 336 Block
E6 Action:T6 Using:T39 Site:T41
T7 Action 427 430 Add
E7 Action:T7 Acts-on:T44
T8 Action 511 519 Incubate
E8 Action:T8
T9 Action 536 541 Rinse
E9 Action:T9 Count:T48
T10 Action 576 585 Visualize
E10 Action:T10 Using:T52
T11 Reagent 54 70 tris-glycine gel
T12 Reagent 82 89 protein
T13 Reagent 93 97 PVDF
T14 Reagent 106 114 membrane
T15 Amount 118 120 1x
T16 Reagent 121 131 TBST Block
T17 Concentration 135 137 5%
T18 Reagent 138 150 non-fat milk
T19 Reagent 151 155 TBST
T20 Time 160 170 30 minutes
T21 Temperature 174 176 RT
R1 Measure Arg1:T16 Arg2:T15
R2 Measure Arg1:T18 Arg2:T17
R3 Meronym Arg1:T18 Arg2:T19
R4 Measure Arg1:T19 Arg2:T17
R5 Setting Arg1:E3 Arg2:T20
R6 Setting Arg1:E3 Arg2:T21
T22 Reagent 188 196 membrane
T23 Reagent 202 215 anti-FMRP 1C3
T24 Concentration 219 225 1:4000
T25 Temperature 229 231 RT
T26 Time 236 239 1hr
R7 Measure Arg1:T23 Arg2:T24
R8 Setting Arg1:E4 Arg2:T25
R9 Setting Arg1:E4 Arg2:T26
T27 Temperature 250 252 4C
T28 Concentration 256 258 5%
T29 Reagent 259 271 non-fat milk
T30 Concentration 272 274 1x
T31 Reagent 275 279 TBST
R10 Or Arg1:T29 Arg2:T31
R11 Measure Arg1:T31 Arg2:T30
R12 Measure Arg1:T29 Arg2:T28
R13 Or Arg1:T25 Arg2:T27
T32 Reagent 243 246 O/N
R14 Or Arg1:T23 Arg2:T32
T33 Reagent 288 296 membrane
T34 Numerical 297 298 5
T35 Concentration 305 307 1x
T36 Reagent 308 312 TBST
T37 Time 318 328 30 minutes
R15 Measure Arg1:T36 Arg2:T35
R16 Setting Arg1:E5 Arg2:T37
T38 Concentration 342 344 5%
T39 Reagent 345 389 IgG-Free, Protease-Free Bovine Serum Albumin
T40 Concentration 393 395 1x
T41 Reagent 396 400 TBST
T42 Time 405 418 30-45 minutes
T43 Temperature 422 424 RT
R17 Measure Arg1:T39 Arg2:T38
R18 Measure Arg1:T41 Arg2:T40
R19 Setting Arg1:E6 Arg2:T42
R20 Setting Arg1:E6 Arg2:T43
T44 Reagent 431 496 HRP-conjugated goat-anti-mouse IgG, light-chain specific antibody
T45 Concentration 500 508 1:10,000
R21 Setting Arg1:E7 Arg2:T45
T46 Time 524 527 1hr
T47 Temperature 531 533 RT
T48 Numerical 542 543 5
T49 Concentration 550 552 1x
T50 Reagent 553 557 TBST
T51 Time 563 573 30 minutes
T52 Reagent 591 603 ECL solution