SKU: 47002108700

UniOne® TR-FRET Human CDK2/CRBN PROTAC Binding Kit

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Description

UniOne® TR-FRET Human CDK2/CRBN PROTAC Binding KitProduct Specification Host Human Stability & Storage 80 Background The assay kit employs homogeneous time resolved fluorescence (TR FRET) technology to measure the interaction between the molecular glue CDK2 Degrader7 (or test compounds) and the DDB1 CRBN complex with CDK2. This method enables simple and rapid high throughput screening of small molecules capable of mediating the interaction between the DDB1 CRBN complex and CDK2. As shown in the

Product Specification


Host Human
Stability & Storage

-80℃

Background

The assay kit employs homogeneous time-resolved fluorescence (TR-FRET) technology to measure the interaction between the molecular glue CDK2 Degrader7 (or test compounds) and the DDB1/CRBN complex with CDK2. This method enables simple and rapid high-throughput screening of small molecules capable of mediating the interaction between the DDB1/CRBN complex and CDK2.

As shown in the figure below, the interaction between DDB1/CRBN and CDK2 is detected using a Eu-labeled anti-Tag1 antibody (TR-FRET donor) and an Ac-labeled anti-Tag2 antibody (TR-FRET acceptor). The molecular glue CDK2 Degrader7 mediates the interaction between DDB1/CRBN and CDK2, bringing the donor and acceptor antibodies into proximity. Excitation of the donor antibody triggers fluorescence resonance energy transfer (FRET) to the acceptor antibody, resulting in a specific emission signal at 665 nm. This signal is proportional to the extent of interaction mediated by CDK2 Degrader7 between DDB1/CRBN and CDK2. The homogeneous assay is straightforward and requires no washing steps.

Components

Product Components and Storage Conditions】

Component

Concentration

100T

500T

2500T

10000T

Storage Temperature

Tag1-CRBN protein

10

5μL

20μL

100μL

400μL

-80℃

Tag2-CDK2 protein

100×

5μL

20μL

100μL

400μL

-80℃

CDK2 Degrader 7

1mM

5μL

5μL

25μL

100μL

-80℃

Anti-Tag1 Eu antibody

5

5μL

50μL

250μL

1mL

-80℃

Anti-Tag2 Ac antibody

100×

5μL

25μL

125μL

500μL

-80℃

Detection buffer

10×

400μL

2mL

10mL

40mL

-80℃


Note: All components should be aliquoted immediately after the first thaw and stored at the recommended temperature to avoid dilution storage and repeated freeze-thaw cycles.

Protocol

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1. Reagent Preparation

1.1 Thaw all reagents to room temperature before use (equilibrate for at least 30 min). The reaction system for 384-well shallow plates is 20 μL (reagent volumes are shown in the table below). Calculate the required volume before preparation. The following preparation is for reference only, using 500 reactions as an example.

Table 1. Reagent Preparation

Reagent Name

Preparation

Volume per Well (μL)

Detection buffer

Add 2 mL of 10× Detection buffer to 18 mL of deionized water, dilute to 1×, and mix well.

-

CDK2 Degrader7

Dilute the compound to the target concentration with 1× Detection buffer according to the reaction system.

2

Tag1-CRBN protein

Take 20 μL of Tag1-CRBN protein stock solution, dilute to 2 mL with 1× Detection buffer, and mix well.

4

Tag2-CDK2 protein

Take 20 μL of Tag2-CDK2 protein stock solution, dilute to 2 mL with 1× Detection buffer, and mix well.

4

Antibody Mix

Take 50 μL of Anti-Tag1 Eu antibody stock solution, add 2.450 mL of 1× Detection buffer, mix well; take 25 μL of Anti-Tag2 Ac antibody stock solution, add 2.475 mL of 1× Detection buffer, mix well; mix the two 1:1 to prepare the Antibody Mix.

10


1.2 Gradient Dilution of Test Samples

Using CDK2 Degrader7 as an example, the diluent is 1× Detection buffer. To minimize matrix interference, it is recommended to use a solution with the same matrix as the test sample for dilution. Adjust the dilution based on the actual concentration of the sample.

Table 2. Gradient Dilution of CDK2 Degrader7 (adjust as needed)

CDK2 Degrader7

Final Concentration (nM)

CDK2 Degrader7 Preparation Concentration (nM)

Preparation Method

1000

10000

1 μL of 1 mM CDK2 Degrader7 + 99 μL of 1× Detection buffer

333.3333

3333.333

20 μL of ① + 40 μL of 1× Detection buffer

111.1111

1111.111

20 μL of ② + 40 μL of 1× Detection buffer

37.0370

370.370

20 μL of ③ + 40 μL of 1× Detection buffer

12.3457

123.457

20 μL of ④ + 40 μL of 1× Detection buffer

4.1152

41.152极u>

20 μL of ⑤ + 40 μL of 1× Detection buffer

1.3717

13.717

20 μL of ⑥ + 40 μL of 1× Detection buffer

0.4572

4.572

20 μL of ⑦ + 40 μL of 1× Detection buffer

0.1524

1.524

20 μL of ⑧ + 40 μL of 1× Detection buffer

Blank

0

0

40 μL of 1× Detection buffer


2. Sample Loading and Controls

2.1 Test samples: Add 4 μL of Tag1-CRBN protein working solution, 4 μL of Tag2-CDK2 protein working solution, 2 μL of gradient-diluted test sample, and 10 μL of mixed Antibody Mix sequentially into the 384-well shallow plate.

2.2 Positive control standard curve: Add 4 μL of Tag1-CRBN protein working solution, 4 μL of Tag2-CDK2 protein working solution, 2 μL of gradient-diluted CDK2 Degrader7, and 10 μL of Antibody Mix.

2.3 Blank control wells: Replace the test sample with 2 μL of 1× Detection buffer.

2.4 NC: Add 10 μL of 1× Detection buffer and 10 μL of Antibody Mix.

After all samples are loaded, centrifuge, seal with plate-sealing film, and incubate at room temperature for 2 hours.

Positive Control Standard Curve

Test Samples

Blank Control Wells

NC

2 μL

Gradient-diluted CDK2 Degrader7

2 μL

Gradient-diluted test sample

2 μL

1× Detection buffer

10 μL of 1× Detection buffer

plus 10 μL of Antibody Mix

4 μL of Tag1-CRBN protein

4 μL of Tag2-CDK2 protein

10 μL of Antibody Mix

Seal the plate with sealing film, and incubate at room temperature for 2 hours.


3. Detection

Measure using a TR-FRET-compatible microplate reader. Excitation wavelength is 320/340 nm, and emission wavelengths are 620 nm and 665 nm.

[Result Calculation]

1) Calculate the signal value (Ratio): Divide the 665 nm fluorescence signal by the 620 nm fluorescence signal and multiply by 10,000.

Ratio = (665/620) × 10,000

2) Calculate the Net signal based on the signal value:

Net signal = (Std - NC)/NC × 100

3) Calculate CV (%):

CV (%) = Standard Deviation/Mean Ratio × 100%

[Data Example]

The following data cannot replace experimental results and is provided as an example only. Results may vary depending on the plate reader used.  

Note: Recommended microplate (384-well plate, white, shallow wells).

```
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Very good bars of soap collection.
My wife doesn’t like the liquid soap because a lot of them say detergent and she says she’s not a dish or a piece of clothing. So this set looks really appealing with a variety of fragrances and scents. It will last you a while. The size of each soap bar is the average size of soap bar. It lathers very well, while being soft on the skin and ganking dirt and odors like it does it’s basically a steal.
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Natural, Gentle, and Smells Great
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I picked these up to try something more natural, and I love them. They lather well, feel gentle on the skin, and leave it feeling clean without drying it out. The scents are really nice too. They feel like good quality bars.
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Kailyn
Massapequa, US
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Longevity, scent, quality 10/10
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I’ve gone through just about the whole box and I will be buying more. I bought these early November 2025 and now that it’s almost May of 2026 I’m down to my last two bars so id say they last awhile. I use them for body wash and will be using them for hand soap once I go through my stash of bar soaps from other brands. It lathers nicely, with a light scent, natural not over bearing chemical scents. I use this as body wash and hand soap. I haven’t noticed any skin irritation, just a good ole natural soap (: I love how there’s no plastic in sight other than maybe amazon packaging
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Better than expected...
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I love going to a craft show or a place that sells a handcrafted soap. A good craftsperson can produce a high-quality soap that cleanses, moisturizes, and leaves a good scent. I gave these a shot after running out of those soaps. They did beat my expectations. I thought these would be a factory produced and chemically. While you can tell these are churned out pretty regularly in some sort of factory, the overall quality is good. Each scent is full and unique without being overpowering. They seem to be natural and not generic chemical smells. Really enjoy each of the unique scents. The bars themselves aren't that big. They come in a piece of cardboard and are very uniform. There is some texture to the soap, as well as an added bit of something to give it some grit. I like it. This isn't lava soap but there is something (about the size of fresh ground pepped) mixed throughout each bar. A drawback overall is how long they last. I would estimate less than a week for a daily user. I thought, given the denseness, they would last for a longer than they do. I'll probably use the entire box in under a month. Of note is the lack of slimy or slipperiness to the soap. I like this. Some soaps that I've used in the past leave you slimy and you can barely hang on to the bar. This rinses very well and doesn't make you feel slimy. Overall, these are a good replacement for the 'true' homemade soap. I am happy with them and would purchase them again.
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