Found inside – Page 52The column was washed with Tris - buffered saline with tween ( TBST : 0.5M NaCl , 0.1 % tween 20 , 100 mM Tris - HCl , pH 8.0 ) and eluted with 4 M Guanidine HCI , 10 mM Tris ( pH 8.0 ) . Elution fractions containing rat BGP were loaded onto ... Found inside – Page 50The strips were soaked in løml tris buffered saline containing ImM EDTA and 0.1 % Tween 20 ( Tris buffered saline / EDTA / Tween- Appendix III ) , for 30 minutes , Fifty ul of 1 mg / ml commercial lectine ( Concanavalin A , Soy Bean Agglutinin ... Found inside – Page 214... Melville , New York ) with a transfer buffer ( 48 mM tris , 39 mM glycine , 20 % methanol , 0.0375 % SDS ; pH 8.3 ) for 1 h ... washed with tris - buffered saline ( TBS ; 20 mM Tris , 500 mM sodium chloride ; pH 7.5 ) containing Tween 20 ( TTBS ... Found inside – Page 43Membranes were then blocked for l h at room temperature in blocking buffer (Tris-buffered saline with 0.1% Tween-20 and 5% Skim milk (SigmaAldrich, ... Found inside – Page 739The curve exhibited a sigmoid with gentle agitation on a platform shaker , and washed thrice shape with a linear part in the range 10-90 % of methylfor 5 min in Tris - Buffered Saline Tween - 20 ( TBST ) ( 50 mM Tris ated DNA . From this we ... Found inside – Page 1780... vehicle ( DMSO ) or phosphate - buffered saline ( PBS ) , DBZ ( 20 basis ... system in 5 % nonfat dry milk in Tris - buffered saline Tween - 20 ( TBST ) ... Found inside... single - stranded DNA binding protein SV40 simian virus 40 TBS tris buffered saline TBST tris buffered saline + 0.05 % Tween 20 TX - 100 triton X - 100 CHAPTER I LITERATURE REVIEW THB MITOCHONDRION The mitochondrion is a xvii. Found inside – Page 524Twenty - four hours after transfection , cells were harvested by trypsinisation and centrifuged at 1000 r / min ( 106g ) for 5 ... fat dry milk in the Trisbuffered saline Tween - 20 ( TBST ) ( 137 mmol / L NaCl , 20 mmol / L Tris , pH 7.6 , 0.1 % Tween ... Found inside – Page 232PLA buffer: 1 mM D-biotin, 0.1% purified bovine serum albumin (BSA), 0.05% Tween-20, 100 nM Goat serum IgG, ... Tris Buffered Saline with Tween-20 ... Found inside – Page 690nonspecific antisera binding , blots were incubated in suggesting high specificity of ligand Hybridization was carried out for 16 hours with the Tris - buffered saline plus Tween ( 20 mM Tris , 154 requisite cDNA probe . Washes and exposure ... Found inside – Page 158The resulting powders were dissolved in 20 mM Tris buffer ( pH 7.7 ) that contained 2 mM DTT , 1 mM EDTA , and 1 mm ... was blocked for 1 h in 3.5 % BSA that was dissolved in Tris - buffered saline containing 0.05 % ( v / v ) Tween 20. Found inside – Page 61Tris-buffered saline with Tween 20 (TBST). Dilute 2 mL of Tween 20 (0.2% v/v) in 0.98 L of Tris-buffered saline, pH 7.4. 2.5 Equipment 1. 2. Found inside – Page 82Approximately 1.0 x106 cpm of probe / ml of hybridization buffer was added and hybridized for 15 hours at 65 ° C ... non - fat powdered milk ( 1 : 1 ) in Tris buffered saline / Tween ( 20 mM TrisHCl pH 8.0 , 152 mM NaCl , 0.05 % Tween - 20 ) . Found inside – Page 30nitrocellulose sheets for 1 hour at 37 ° C with gentle shaking in Tris buffered saline ( TBS ; 20 mM Tris , 0.9 % NaCl , 0.05 % Tween -20 , 1 : 10,000 merthiolate , pH 7.5 ) containing 3 % gelatin . The sheets were then washed 3 times in TBST ... Found inside – Page 51Following three 10 min washes in tris - buffered saline containing Tween 20 ( TBS - T ) , membranes were incubated with the caspase - 3 antibody ( Cell Signaling , Beverly , MA , USA ) ( 1 : 750 in TBS - T plus 3 % BSA ) overnight at 5 ° C ... Found inside – Page 353380 or CAN99-81 ) ( 1 : 2,000 diluted in 5 % skim milk ) for 1 h at room temperature , washed with Tris - buffered saline containing 0.2 % Triton X - 100 and 0.05 % Tween 20 , and incubated with goat anti - guinea pig antibody conjugated with ... Found inside – Page 276The blotted membranes were placed in blocking buffer ( Tris - buffered saline , pH 8.0 , supplemented with 0.3 % Tween 20 and 5 % nonfat dry milk ) for 1 h ... Found inside – Page 179PBS-T: PBS, 0.05% Tween-20. 23. TBS-T: Tris-buffered saline 0.5% Tween-20. 23. Wash buffer 1: 10 mM Tris-HCl, pH 7.4; 1 mM EDTA; 150 mM NaCl; ... Found inside – Page 32110X CP Ligation buffer: 1 M Tris-HCl, pH 8.5, 100 mM MESNA (Sigma) (see Note 7). ... Tris-buffered saline with Tween-20 and Triton X-100 (TBSTT): 20 mM ... Found inside – Page 99Equilibrate the gels with transfer buffer and blot the minigels onto NC ... Elution, and Storage Tris-buffered saline with Tween-20 (TBST) (10 mM Tris, ... Found inside – Page 109The membrane was blocked with blocking buffer , ( 0 . 5 % casein ( w / v ) in Tris buffered saline ( TBS ) , pH 7 . 5 ) with 0 . 03 % Tween - 20 ( v / v ) . Immunodetection All immunodetection steps were performed at room temperature , ~ 23°C ... Found inside – Page 20The PVDF membrane was blocked with 5% non-fat dry milk/Tris-buffered saline (pH 7.5) plus 0.1% Tween 20 (TBST) solution and subjected to the addition of Dlx-3 primary antibody (1° AB from Dr. Mark Roberson's laboratory-Ithaca, NY) and ... Found inside – Page 20Materials and Methods Buffers Reassembly Buffer : 10 mM MES ( 2 - Nmorpholinoethane sulfonic acid ) at pH 6 . 7 , 150 mm ... Phosphate Buffered Saline ( PBS ) : 0 . 14 M sodium ... Tris Tween - 20 Buffered Saline ( TTBS ) : 0 . 5 M sodium ... Found inside – Page 1902After immunoprecipitation , beads were washed thrice with MDB buffer ( 50 mM Tris , pH 7 . ... and the membrane was blocked with 3 % BSA in Tris - buffered saline / Tween 20 and incubated with antibodies before development with ECL . Found inside – Page 74The buffer solution used during the transfer contained 20 % ( v / v ) methanol , a generic formulation for low molecular ... The blot was washed for 5 minutes 3 times in a TRIS buffered saline solution containing 0.05 % TWEEN 20 ( TBST ) to ... Found inside – Page 388Reagents TBST (Tris-buffered saline with Tween 20): 50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 0.05% (v/v) Tween 20 BLOTTO" [5% (w/v) powdered milk in phosphate-buffered saline]: 5% (w/v) dry milk (Carnation brand, nonfat milk), 0.01 M ... Found inside – Page 312Tris Buffered Saline with Tween 20 ( TTBS ) : ( 1x TBS + 0.05 % v / v Tween - 20 ( Sigma , cat . no . P - 9416 ) ) . This buffer is usually made up fresh ... Found inside – Page 264Running buffer* 1.5% (w/v) Tris base, pH 8.3 15 g 7.2% (w/v) Glycine 72 g ... water Tris buffered saline with Tween 20 (TTBS)** 0.5 ml Tween 20 to 1 l of ... Found inside – Page 86The membrane was then blocked overnight at 4 C in TBST ( tris buffered saline + 0.05 % Tween - 20 ) + 5 % nonfat dry milk . The membrane was then blocked an additional 45 minutes at room temperature in SuperBlock ( 0.2 M glucose , 10 ... Found inside – Page 128A typical 1X transcription buffer may contain 40 mM Tris-HCl, pH 7.5; ... 1X PBST (phosphate-buffered saline with Tween-20:Sigma, St. Louis, MO; cat. no. Found inside – Page 1959... a random sample of 26 of the nitrocellulose sheets that had been placed over urine samples during behavioural tests were probed with MUP antisera after blocking with 0.5 % ( w / v ) Tween 20 in Tris buffered saline ( TBS ) pH 7.5 for 1 h . Found inside – Page 29Tris Buffered saline (TBS): The TBS buffer consists of 50 mM Tris-Cl, ... Tween-20 solution • TBS-Tween (0.1%): Add 1 ml of Tween-20 solution to 999 ml of ... Found inside – Page 244TRIS-buffered saline, pH 7.6 (usually prepare as 10× stock). ... Make on day of use (1 L for two gels): ○ TRIS-buffered saline ○ 0.1 % Tween-20. Found inside – Page 7314Briefly, the membranes were blocked with 5% skimmed milk in Tris-buffered saline containing 0. 1 % Tween 20 and incubated overnight at 4°C with the primary antibodies diluted in this buffer before proceeding to the alkaline phosphatase ... Found inside – Page xii... TCR TE TFA TLCK TPCK Tris TTBS UTP VVA WGA ZAPNA N-hydroxysuccinimide ... Tris-buffered saline containing Tween 20 T cell receptor TriS-EDTA buffer ... Found inside – Page 110... to - side shaker Buffers - Tris - buffered saline ( TBS , pH 7.6 ) : - 10 mM Tris / HCl - 150 mM NaCl Tris - buffered saline , 0.2 % Tween 20 ( TBS - T ) ... Found inside – Page 35Membranes were blocked 0.512 hr in 3 % ( w / v ) dry milk , 0.1 % ( v / v ) Tween - 20 / Tris buffered saline ( TBS ) followed by an incubation with a 1 : 3000 dilution of the affinity purified antibody ( Dinah ) in 1 % dry milk , 0.1 % Tween - 20 / TBS ... Found inside – Page 650Chemicals All purified enzymes , coenzymes , substrates , and buffers were purchased from Sigma Chemicals ... Tris - buffered saline ( TBS ) for 15 min , and blocked with 1 % bovine serum albumin ( fraction V ) and 0.5 % Tween 20 for 1 h . Found inside – Page 13The addition of 1 mL homogenizing buffer ( 0.25 mol / L sucrose , 10 mmol / L Tris - HCl , and 0.145 mol / L NaCl ( pH 7.4 ) ... of MLCL AT was examined by immunoblotting in 10 mL Tris - buffered saline containing 0.1 % Tween - 20 with antiFig . Found inside – Page 252After a 1 - h transfer , the membranes were washed in Tris - buffered saline with Tween 20 ( TBST ) ( 24.2 g Tris , 80.0 g NaCl , 0.1 % Tween 20 in 10L , pH 7.6 ) . The membranes were then blocked by gently agitating them in TBST with 5 ... Found inside – Page 113Sample buffer ( 50 ul ; 5X buffer containing 0.25M Tris - HCI pH 6.8 , 50 % glycerol , 10 ... After washing with TBST buffer ( BupH Tris buffered saline containing 0.1 % Tween 20 ) , immunodetection was performed with either anti - Bak ( H - 211 ... Found inside – Page 20Twenty micrograms of proteins were resolved by 10% SDS-PAGE and transferred to PVDF ... After three times washing with Tris buffered saline Tween20 (TBST), ... Found inside – Page 238Proteins were resolved by 12 % SDS - PAGE and transferred to nitrocellulose membranes ( BA 85 , Schleicher & Schuell ) . The membranes were blocked with 5 % BSA in Tris - buffered saline / Tween 20 ( TBST ) ( 10 mM Tris - HCl , pH 7.6 ... Found inside – Page 81buffer ( 25 mM Tris , pH 7.5 , 100 mM NaCl , 0.5 % ( v / v ) Nonidet P - 40 [ NP - 40 ] , 30 ug E - 64 ( Boehringer - Mannheim ) ... The blots were incubated for 1 hour in blocking buffer ( Tris - buffered saline ( TBS ] , 5 % NP - 40 , 1 % Tween 20 , 1 ... Found inside – Page 6797Membranes were then washed twice ( 10 - min machine wash ) in Tris - buffered saline - Tween ( 20 mM TrisHCI ( pH 7.5 ) , 150 mM NaCl , and 0.05 % Tween 20 ) . Membranes were then incubated at room temperature with a 1/5000 dilution ... Found inside – Page 49P Add 200 pL of Tris-buffered saline containing 0.1% gelatin and 0.05% Tween 20 to each well to block the remaining binding sites on the plate surfaces and ... Found inside – Page 34... Buffers Tris-buffered saline (TBS): 20 mM Tris, 500 mM NaCl, pH 7.5 Tris-buffered saline-Tween (TBS-T): TBS, containing 0.05% Tween 20 Blocking buffer: ... Found inside – Page 80... Appendix 1) Skim milk (See Appendix 1) Tris-buffered saline with Tween-20® (See Appendix 1) ... Then soak the membrane in 1× transfer buffer for 1 min. Found inside – Page 100Embryos were washed in TBS ( tris - buffered saline ) -0.1 % Tween - 20 ( TBST ) , blocked in TBST with 0.5 % Triton - X - 100 and 5 % heat - inactivated goat serum for 2 hours at room temperature , and incubated in primary antibody ( HNK - 1 ...
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