Common Solutions

MOPS NUPAGE RUNNING BUFFER: 104,6g MOPS, 60,6 Tris Base, 10g SDS, 3 g EDTA in 500ml final(50mM MOPS,50mM Tris,0,1%SDS,1mM EDTA) pH 7.7 do not use acid or base to adjust the pH.

BLU DI COOMASSIE 125 ml di isopropanol, 30 ml acetic acid, 325 ml H20, 1,25 gr Brillant Blue. Dissolve O.N. Stirring and finally filter on paper.

DESTAIN / FIXING SOLUTION 10% acetic acid, 10% methanol

EDTA 0.5M pH 8.0: dissolve186.1 g EDTA disodic salt in H₂0. Stirr and add NaOH till pH=8.5 add water till11. Pay attention: EDTA will dissolve only if pH= 8

NaCl 5M: dissolve 292.2 g NaCl in H20 in 1L final water. Stock RT

LOADING BUFFER: 5 ml SAMPLE BUFFER 4X, 4 ml SDS 20%, BpB powder

PBS 10X: To make 1 L of 10X PBS stock solution, combine 17.8 g of Na2HPO4 (FW 177.99) , 2.4 g of KH2PO4 (FW 136.09) , 80 g of NaCl (FW 58.44), 2 g of KCl (FW74.55). Adjust final volume to 1 L. The pH of the 10X stock will be approximately 6.8, but when diluted to 1x PBS the pH should change to 7.4. If necessary, the pH can be adjusted using hydrochloric acid or sodium hydroxide.

(OLD) PBS 10X: prepare following solution: NaH₂P04 1M pH 4.0 (137,99 g/mol); Na₂HP0₄ 1M pH 9.0 (177,99 g/mol hard to dissolve) mix solution starting from pH 9 solution till pH 7.3. From buffer pH 7.3 take 50ml and add 45g NaCl and H20 to 500ml final.

SAMPLE BUFFER 4X 5,25 ml UPPER UFFER + 20 ml glycerol 100%

SDS (20% w/v): Dissolve 20 g SDS in 100 ml H₂0 at 60°C.

STN (washing beads in pulse chase experiment) 10mM TRIS HC1 pH:7.0, 0.25% NP40, 150mM NaCl

H20 lavaggi (last wash for beads in pulse chase experiment) 5mM TRIS HCl pH:7,0

STRIPPING SOLUTION 62,5mM Tris HCl pH=6,7, add beta mercaptoethanol (100 mM final); NaOH

TAE 50X (pH 8 ):Tris base 242 g, EDTA pH 8 100ml 0.5 M, Acido acetico glaciale 57.1 ml up to 1L with water.

TAMPONE DI CORSA TRIS GLICINA 10X: dissolve 30g di tris a di 144g glicina in 11 di H₂O . Add final 0.1% SDS before use. (4 litri: 120 g Tris; 576 g glicina)

NP40 Lysis Buffer: 150mM NaCI, 10mM TRIS (pH. 7. 5), 1% NP40

TRANSFER BUFFER 25 mM Tris base, 192 mMglicina,20% metanolo Sciogliere 3,03 gr di Tris a 14,4 gr di glicina in circa 750 ml di H20; aggiungere 200 ml di metanolo a portare a volume finale di 1 1

TRIS GLICINA (10X) per RUNNING BUFFER SDS PAGE 144g GLICINA, 30g TRIS BASE bring to 1L con H₂O or 576g GLICINA, 120g TRIS BASE bring to 4L with H₂O

TRIS HC1 pH 6.8 (0.5M): Dissolve 6 g Tris in 40 ml H₂O, adjust pH with 48 ml HCl 1M to 100 ml final with H₂O.

TRIS HCI IM pH 7.5: dissolve 121.1 g TRIS base in H₂O. Adjust pH with HCI (65ml pH 7.5) bring to 1L.

TRIS HCL pH 8.8 (3M): Dissolver 36.3 g tris in 40 ml di H₂O, adjust pH with 48 ml HCI 1M bring to 100 ml with H₂O

UPPER GEL BUFFER 4X 0, 5 M Tris HCI, 0, 4 % SDS, pH 6, 8. Add 30,3g Tris base, 10 ml SDS 20% pH 6.8 with HCI add H₂O to 500 ml final

LOWER GEL BUFFER 4X 1,5 M Trs HC1, 0,4 % SDS, a pH 8,8. Add 90,9g Tris base, 10 ml SDS 20% pH 8.8 with HCI add H₂O to 500 ml final