RBC Lysis Buffer, TBST, NP-40 Cell Lysis Buffer, Proteinase K Solution,
RIPA buffer, Modified RIPA buffer, ELISA coating buffer
90g NH4Cl (0.1555M)
10g KHCO3 (0.01M)
370mg EDTA (0.1mM)
Or 1X RBC lysis buffer:
To 800ml dH2O add:
8.3g ammonium chloride (NH4Cl )
1.0g potassium bicarbonate (KHCO3)
1.8ml of 5% EDTA
Filter sterilize through 0.2um filter.
Bring to final volume of 1000ml with dH2O.
(50 mM Tris, 150 mM NaCl, 0.05% Tween-20, pH 7.4)
For 1 liter of 1X Tris-buffered saline with Tween-20 (1X TBST buffer)
-Dissolve in 800ml of distilled H2O:
8g NaCl
0.2g KCl
3g Tris base
-Add 500ul Tween-20
-Adjust the pH to 7.4 with HCl
-Add distilled H2O to 1L
For 1 liter of 10X Tris-buffered saline with Tween-20 (10X TBST buffer)
-Dissolve in 800ml of distilled H2O:
80g NaCl
20g KCl
30g Tris base
-Add 500ul Tween-20
-Adjust the pH to 7.4 with HCl
-Add distilled H2O to 1L
50mM Tris-HCl pH 8.0
150mM NaCl
1% NP-40
plus protease inhibitors: Aprotinin, leupeptin, pepstatin: 1ug/ml each
Add 1mM PMSF immediately before use.
See: P9070, Protease Inhibitor Cocktail
(20 ug/ml in TE Buffer, pH 8.0):
TE Buffer (50mM Tris Base, 1mM EDTA, 0.5% Triton X-100, pH 8.0):
Tris Base -------------------------------- 6.10 g
EDTA ------------------------------------- 0.37 g
Triton X-100 ---------------------------- 5 ml
Distilled water ------------------------- 1000 ml
Mix to dissolve. Adjust pH 8.0 using concentrated HCl (10N HCl). Store at room temperature.
50mM Tris, pH 8.0
150mM sodium chloride
0.1% SDS
0.5% Sodium deoxycholate
1% NP40
plus protease inhibitors: Aprotinin, leupeptin, pepstatin: 1ug/ml each
Add 1mM PMSF immediately before use.
See: R2031-75, RIPA Lysis Buffer
P9070 , Protease Inhibitor Cocktail
10 mM Tris, pH 7.4
100 mM NaCl
1 mM EDTA
1 mM EGTA
1 mM NaF
20 mM Na4P2O7
2 mM Na3VO4
1% Triton X-100
10% glycerol
0.1% SDS
0.5% deoxycholate
plus protease inhibitors: Aprotinin, leupeptin, pepstatin: 1ug/ml each
Add 1mM PMSF immediately before use.
See: C2605-70, Cell Extraction Buffer
P9070, Protease Inhibitor Cocktail
To 900ml dH20, add:
5.3g of Na2CO3
4.2g of NaHCO3
1g sodium azide
Adjust pH to 9.6
Adjust volume to 1L with additional dH2O
Note: For coating step add 0.1-1ug/ml of coating antibody to buffer
and incubate at 4°C for 12-24 hours.
Orange G for DNA gels
Combine:
3.9mL Glycerol
0.5mL 10% SDS
0.2mL 0.5M EDTA
10mg Orange G
Bring mixture to 10mL with ddH2O.
Aliquot in 1.5mL tubes and store at -20C.
From https://med.wmich.edu/sites/default/files/Pioli_Lab_10X_DNA_Loading_Dye_Recipe.pdf