30% (w/v) acrylamide solution
Acrylamide 29g
Methylene bisacrylamide 1g
Add deionized water, dilute to 100 ml, adjust the pH of the solution to less than 7.0, store in a brown bottle at room temperature or 4 °C.
10% SDS
SDS 10g
Distilled water to 100ml
Dissolve in a 50 ° C water bath and store at room temperature. If precipitation occurs during long-term storage, it can still be used after the water bath has melted.
1.5mol/L Tris·HCl (pH8.8)
Tris (MW121.14) 45.43g
Ultra pure water 200ml
After dissolving, adjust the pH to 8.8 with concentrated hydrochloric acid, and finally make up to 250 ml with ultrapure water and store at room temperature.
0.5 mol/L Tris·HCl (pH 6.8)
Tris (MW121.14) 15.14g
Ultra pure water 200ml
After dissolving, adjust the pH to 6.8 with concentrated hydrochloric acid, and finally make up to 250 ml with ultrapure water and store at room temperature.
10% ammonium persulfate (AP)
Ammonium persulfate  0.1g
Ultrapure water 1.0ml
After dissolution, it was stored at 4 ° C in the dark and stored for 1 week. (You can first put the persulfate amine dry powder into the EP tube, put a few tubes at a time, add ultrapure water when using)
20% Tween20
Tween20 20ml
Distilled water to 100ml
After mixing, store at 4 °C.
1.74mg/ml (10mmol/L) PMSF
PMSF 0.174g
Isopropyl alcohol 100ml
After dissolving, it was dispensed in a 1.5 ml centrifuge tube and stored at -20 °C.
Single detergent lysate (PMSF)
1mol/L Tris·HCl (pH8.0) 2.5ml
NaCl 0.438g
TritonX-100 0.5ml
Distilled water to 50ml
After mixing, store at 4 °C. At the time of use, PMSF was added to a final concentration of 100 μg/ml (0.87 ml of lysate was added to 1.74 mg/ml of PMSF 50 μl).
RIPA Lysis Formula
50 mM Tris-HCl pH 7.4
150 mM NaCl
1 mM PMSF (plus before use)
1 mM EDTA
1% Triton x-100
1% sodium deoxycholate
0.1% SDS
G250 Coomassie Brilliant Blue Solution (for protein quantification)
Coomassie Brilliant Blue G250 100mg
95% ethanol 50ml
Phosphoric acid 100ml
Distilled water to 1000ml
When preparing, first dissolve the Coomassie blue dye with ethanol, add phosphoric acid and water, mix well, filter with filter paper, and store at 4 °C.
100mg/ml bovine serum albumin (BSA)
BSA 0.1g
0.15 mol/L NaCl 1ml
Store at -20 ° C after dissolution. When preparing the protein standard curve, it was diluted 100-fold with 0.15 mol/L NaCl to 1 mg/ml and stored at -20 °C.
Reduced 5X SDS Loading Buffer Â
0.5 mol/L Tris·HCl (pH 6.8) 2.5 ml
Dithiolitol (DTT, MW154.5) 0.39g
SDS 0.5g
Bromophenol blue 0.025g
Glycerin 2.5ml
After mixing, dispense in a 1.5 ml centrifuge tube and store at 4 °C.
Electrophoresis fluid buffer Â
Tris (MW121.14) 3.03g
Glycine (MW75.07) 18.77g
SDS 1g
Distilled water to 1000ml
After dissolution, it can be stored at room temperature, and the secondary solution can be reused 3 to 5 times.
(The electrophoresis fluid can be recycled and reused. Generally, the recovered electrophoresis fluid is added to the lower half of the vertical electrophoresis tank, and the upper half is preferably freshly used.)
(Can be prepared into 10× running buffer for storage and diluted 10 times)
Transfer buffer  Â
Semi-dry
24mmol/L Tris (MW: 121.14) 1.4536 g
192mmol/L glycine (MW: 75.07) 7.20672 g
20% methanol 100 ml
Distilled water to 500ml
Impregnation (wet rotation)
48mmol/L Tris 2.375g
39mmol/L glycine 11.25g
20% methanol 200ml
0.0375% SDS 0.375g
Distilled water to 1000 ml
After dissolving, it can be stored at room temperature. The secondary solution can be reused 3 to 5 times (the solution is best stored in a 4 degree refrigerator, and the maximum use is about 5 times, otherwise the transfer efficiency is affected). (First dissolve the glycine, Tris and SDS with distilled water, then add methanol, and finally make up the liquid. If you add methanol first, it is difficult to dissolve glycine, Tris and SDS)
(It can be prepared as 2× transfer buffer for storage and used after dilution)
10X Li Chunhong dyeing solution
Li Chunhong S 2g
Trichloroacetic acid 30g
Sulfosalicylic acid 30g
Distilled water to 100ml
TBS buffer
1 mol/L Tris·HCl (pH 7.5) 10ml
NaCl 8.8g
Distilled water to 1000ml
(It can be prepared as 10×TBS buffer for dilution and 10 times diluted)
TBST buffer Â
20% Tween20 1.65ml
TBS 700ml
It can be used after mixing, and it is best to use it now.
Blocking solution Â
Skimmed milk powder (domestic) 5g
TBST 100ml
It is best to use it now.
Eluting antibody buffer
14.4 mol/L β-mercaptoethanol 700 μl (ventilated kitchen Riga)
SDS 2g
0.5mol/L Tris·HCl (pH6.8) 12.5ml
Ultrapure water to 100ml
When it is formulated, it is carried out in a ventilated kitchen. Store at 4 ° C. Can be reused once. (can be used)
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