Proteomic pre-treatment kit

Cat. No.: P1456469

GRADE & PURITY BioReagent, Proteomics grade, Sterile

Overview

Product Introduction:
At present, the conventional proteome sample pretreatment process has many problems, such as poor reproducibility, poor method stability, low sensitivity, being very time-consuming, and inability to be automated. To address the above issues, Aladdin has launched a new generation of "All In One Tube" innovative products. This product is mainly designed for the processing of trace proteins (5-100μg). Even researchers without omics background can quickly prepare pretreated samples, which can be used for subsequent mass spectrometry analysis.

Experimental Flowchart of Proteomics Pretreatment Kit (P1456469)

Product Components and Storage Conditions:

Storage Unit No.	Storage Unit Name	Component No.	Component Name	Appearance	12T	24T	48T	Storage
P1456469NT	Proteomic Pretreatment Kit (RT Components)	P1456469D	Stop Buffer	Liquid	0.375 mL	0.75 mL	1.5 mL	Room temperature
		P1456469E	Wash Buffer I	Liquid	5 mL	10 mL	20 mL	Room temperature
		P1456469F	Wash Buffer II	Liquid	3.75 mL	7.5 mL	15 mL	Room temperature
		P1456469G	Wash Buffer III	Liquid	3.75 mL	7.5 mL	15 mL	Room temperature
		P1456469H	Elution Buffer	Liquid	6.25 mL	12.5 mL	25 mL	Room temperature. Store in the dark.
		P1456469I	Loading Buffer	Liquid	0.25 mL	0.5 mL	1 mL	Room temperature
		P1456469J	Desalting Column	/	12 T	24T	48T	Room temperature
P1456469WT	Proteomic Pretreatment Kit (2-8℃ Components)	P1456469C	Balance Buffer	Liquid	3 mL	6 mL	12 mL	2-8℃
P1456469DT	Proteomic Pretreatment Kit (-20℃ Components)	P1456469A	Lysis Buffer	Liquid	1.25 mL	2.5 mL	5 mL	-20℃. Store in the dark.
		P1456469B	Digest	Liquid	0.05 mL	0.1 mL	0.2 mL	-20℃

Important Notes：

Each component is individually packaged according to its storage temperature (Room temperature/2-8 °C/-20 °C) upon shipment, with differentiated temperature-controlled transportation to guarantee product performance. Orders containing materials requiring multiple temperature zones may be delivered in separate batches; please refer to the actual received goods. Upon receipt, store components separately at the temperatures specified in the component list. The shelf-life is 12 months from the date of manufacture.

Product Features:

Trace proteins: Designed for the pretreatment of 5-100μg proteins.

Simple operation: Enables rapid preparation of pretreated samples, requiring only a metal bath and a conventional centrifuge.

High stability: Strict quality inspection for each batch ensures high reproducibility of experimental results.

Operating Procedure:

Lysis
① Add 50μL of Lysis buffer to the EP tube containing the sample and mix by shaking.
② Place the EP tube with Lysis buffer in a 95°C water bath for 10 minutes, then take it out and cool to room temperature.
Enzymatic Digestion
① Add 225μL of Balance buffer to the EP tube that has cooled to room temperature.
② Add 3μL of Digest, mix well, and perform enzymatic digestion with shaking at 37°C and 1200rpm overnight (16 hours is recommended).
Desalting (at room temperature)
① Add 25μL of Stop buffer to the sample and vortex to mix.
② Add 320μL of Wash buffer 1, shake vigorously for 3 minutes, centrifuge at 15000rpm for 3 minutes, and remove the supernatant.
③ Transfer the lower layer sample into the Tip column, centrifuge at 2500rpm for 3-5 minutes until all liquid is centrifuged down. If the liquid flow rate is slow, the speed can be increased.
④ Add 200μL of Wash buffer 2 (shake for 10-20 seconds before use) to the desalting column, centrifuge at 2500rpm for 3-5 minutes until all liquid is centrifuged down.
⑤ Add 200μL of Wash buffer 3 to the desalting column, centrifuge at 2500rpm for 3-5 minutes until all liquid is centrifuged down.
⑥ Put the desalting column into a new EP tube, add 200μL of Elution buffer to the desalting column, centrifuge at 2000rpm for 3-5 minutes until all liquid is centrifuged down.
⑦ Repeat step ⑥, collect the eluates from both times, and freeze-dry them.
⑧ Add 10μL of Loading buffer, vortex vigorously for 3 minutes, centrifuge at 20000g for 10 minutes, take an appropriate amount of sample, and then mass spectrometry detection can be performed. Taking the HF-X instrument as an example, 0.5-1μg of sample is sufficient for loading.

Precautions:
After aliquoting, Digest should be stored at -20°C.
After aliquoting, Lysis Buffer should be stored at -20°C and avoid repeated freezing and thawing.
This product is limited to scientific research use by professionals, and must not be used for clinical diagnosis or treatment, nor for food or drugs.

Specifications

Specifications & Purity

BioReagent, Proteomics grade, sterile

Grade

BioReagent, Proteomics grade, Sterile

Application

Mass Spectrometry Analysis, Proteomic Sample Preparation

Sensitivity

Light-sensitive

Storage and Shipping

Storage

Store at 2-8°C,Protected from light,Room temperature,Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Normal,Wet ice,Ice chest + Ice pads

Stability And Storage

Store at room temperature/2-8℃/-20℃ long term (12 months).

Contents & Storage

Storage Unit No.	Storage Unit Name	Component No.	Component Name	Appearance	12T	24T	48T	Storage
P1456469NT	Proteomic Pretreatment Kit (RT Components)	P1456469D	Stop Buffer	Liquid	0.375 mL	0.75 mL	1.5 mL	Room temperature
		P1456469E	Wash Buffer I	Liquid	5 mL	10 mL	20 mL	Room temperature
		P1456469F	Wash Buffer II	Liquid	3.75 mL	7.5 mL	15 mL	Room temperature
		P1456469G	Wash Buffer III	Liquid	3.75 mL	7.5 mL	15 mL	Room temperature
		P1456469H	Elution Buffer	Liquid	6.25 mL	12.5 mL	25 mL	Room temperature. Store in the dark.
		P1456469I	Loading Buffer	Liquid	0.25 mL	0.5 mL	1 mL	Room temperature
		P1456469J	Desalting Column	/	12 T	24T	48T	Room temperature
P1456469WT	Proteomic Pretreatment Kit (2-8℃ Components)	P1456469C	Balance Buffer	Liquid	3 mL	6 mL	12 mL	2-8℃
P1456469DT	Proteomic Pretreatment Kit (-20℃ Components)	P1456469A	Lysis Buffer	Liquid	1.25 mL	2.5 mL	5 mL	-20℃. Store in the dark.
		P1456469B	Digest	Liquid	0.05 mL	0.1 mL	0.2 mL	-20℃

Important Notes：

Each component is individually packaged according to its storage temperature (Room temperature/2-8 °C/-20 °C) upon shipment, with differentiated temperature-controlled transportation to guarantee product performance. Orders containing materials requiring multiple temperature zones may be delivered in separate batches; please refer to the actual received goods. Upon receipt, store components separately at the temperatures specified in the component list. The shelf-life is 12 months from the date of manufacture.