技术文章

Buffer preparation guide

Buffer formula


1.0 mol/L TrisHCl(pH 6.8)

Tris 30.28 g
Ultrapure water 200 mL

After dissolution, the pH was adjusted to 6.8 with concentrated hydrochloric acid, and finally the volume was made up to 250 mL with ultrapure water. After autoclaving, the solution was stored at room temperature.


1.5mol/L TrisHCl(pH 8.8)

Tris 45.43 g
Ultrapure water 200 mL

After dissolving, adjust the pH to 8.8 with concentrated hydrochloric acid, and finally dilute to 250 mL with ultrapure water, sterilize at a high temperature, and store at room temperature.


10% SDS

Take 10 g of SDS, add distilled water to 100 mL, dissolve in a 50 ℃ water bath, and store at room temperature. If precipitation occurs during long-term storage, it can still be used after dissolving in a water bath.


10% Ammonium persulfate (APS)

Take 0.1 g of ammonium persulfate, dissolve in 1.0 mL of ultrapure water, and store at 4 ℃ for 1 week. (Long-term storage at -20 ℃)


30% Acrylamide

Acrylamide
29 g
N,N'-methylenebisacrylamide
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Dissolve in a total volume of 50 mL of water and make up to 100 mL. Store at room temperature in a brown bottle.


10× PBS(2L)

NaCl 160 g
KCl 4 g
Na2HPO4·12H2O 22.9 g
KH2PO4 4 g
dH20 1 L

NaOH adjusts the pH to 7.4 and brings to a volume of 2 L. Store at room temperature.


Reduction type 5×SDS sample loading buffer

0.5 mol/L TrisHCl(pH6.8) 2.5 mL
Dithiothreitol (DTT) 0.39 g
SDS 0.5 g
Bromophenol blue 0.025 g
Glycerin 2.5 mL

After mixing, aliquot into 1.5 mL centrifuge tubes and store at 4 °C for several weeks or at -20 °C for long-term storage.


Electrophoresis liquid buffer

Tris 3.03 g
Glycine 18.77 g
SDS 1 g

Add distilled water to dissolve and dilute to 1000 mL. Store at room temperature. One solution can be reused 3 to 5 times.


Transfer buffer


1X electrolysis:

Glycine 11.25 g
Tris 2.375 g
SDS 0.375 g
Methanol 200 mL

Add distilled water to dissolve and bring to a final volume of 1000 mL. Store at 4 °C. One solution can be reused 3 to 5 times.


TBS buffer

NaCl 8.5 g
Tirs 1.2 g

Dissolve in distilled water, adjust the pH to 7.2 with glacial acetic acid, and make up to 1000 mL. Store at room temperature.


Elution solution: TBST solution

TBS solution 1000 mL, 1 mL Tween 20, mix well and use immediately.


Block solution (TBST buffer containing 5% skim milk powder)

Skimmed milk powder 5 g
TBST 100 mL

Use after dissolving, for single use only.


RIPA (medium) lysis buffer

Reagent Amount Final concentration
Tris-base 0.6055 g 50 mM
NaCl 0.8766 g 150 mM
EDTA·2Na.2H2O 0.037224 g 1 mM
SDS 0.1 g 0.1%
Sodium deoxycholate 0.5 g 0.5%
NP-40 1 mL 1.0%

Tris-base, NaCl, EDTA, SDS were weighed and dissolved in distilled water, concentrated HCl was used to adjust the pH to 7.4, and 10% sodium deoxycholate was prepared first. 5 mL was added to make it 0.5%, and then 1 mL NP-40 was added. After mixing, the volume was made up to 100 mL, divided and stored at -20°C.


For more product details, please visit Aladdin Scientific website.

https://www.aladdinsci.com/

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Da — 若无特别说明,分子量单位默认为道尔顿。   Mw — 重均分子量。   Mn — 数均分子量。

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阿拉丁科学.《Buffer preparation guide》. 阿拉丁知识库,更新于 2024年10月24日。 https://www.aladdin-e.com/zh_cn/faqs/buffer-preparation-guide-en.html
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