SKU: 85937761780

UniOne® TR-FRET Human VCB(VHL Complex)Binding Kit

Sale price$328.50 Regular price$365.00
Save 10%

Pay in installments of $91.25 with ShopPay, AfterPay and Klarna

Shipping Estimate
USA
  • USA
  • CAN

Ships within 48 hours · Estimated delivery Aug 27 - Sep 1

Promo Codes Available:

For Your Every Summer RSVP, with Code: SUMMER15

Description

UniOne® TR-FRET Human VCB(VHL Complex)Binding KitProduct Specification Host Human Stability & Storage 80 Background The reagent kit employs homogeneous time resolved fluorescence technology (TR FRET) for measuring the interaction between Human VCB (VHL Elongin C Elongin B Complex) and Human HIF 1. The Tag2 Tracer is a fluorescently labeled HIF 1 peptide fragment. This method enables simple and rapid high throughput screening of inhibitors and antibody blockers. As shown in the figure, the

Product Specification


Host Human
Stability & Storage

-80℃

Background

The reagent kit employs homogeneous time-resolved fluorescence technology (TR-FRET) for measuring the interaction between Human VCB (VHL/Elongin-C/Elongin-B Complex) and Human HIF-1α. The Tag2-Tracer is a fluorescently labeled HIF-1α peptide fragment. This method enables simple and rapid high-throughput screening of inhibitors and antibody blockers.

As shown in the figure, the interaction between VCB and Tracer is detected using a Eu-labeled anti-Tag1 antibody (TR-FRET donor) and an Acceptor-labeled SA (TR-FRET acceptor). Since the binding of VCB and Tracer brings the donor antibody and acceptor antibody 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. VH032 can block the binding of VCB and Tracer, preventing the generation of FRET signals. The stronger the blocking effect of the screened drug on the VCB-Tracer interaction, the lower the signal. The signal is proportional to the degree of interaction between VCB and Tracer. No washing steps are required.

Components

组分

浓度

100T

500T

2500T

10000T

储存温度

Tag1-VCB protein

100 ×

5μL

20μL

100μL

400μL

-80℃

Tag2-Tracer

100 ×

5μL

20μL

100μL

400μL

-80℃

Eu-anti-Tag1

50 ×

10μL

50μL

250μL

1000μL

-80℃

SA-Ac

100 ×

5μL

25μL

125μL

500μL

-80℃

Detection buffer

1 ×

3mL

14mL

70mL

240mL

-80℃


Protocol

1. Reagent Preparation

1.1 Before use, thaw all reagents at room temperature (room temperature equilibration for at least 30 min). The 384-well shallow well plate reaction system volume is 20 μL (reagent volumes for the reaction system are shown in the table); calculate the required volume before preparation and prepare as needed; the following preparation is for reference only, taking 500 times as an example.

Table 1. Reagent Preparation

Reagent Name

Preparation

Volume per Detection Well μL

Tag1-VCB protein

Take 20 μL 100 × Tag1-VCB protein stock solution, add to 1.98 mL 1×Detection buffer to dilute to 1×, mix and set aside.

4μL

Tag2-Tracer

Take 20 μL 100 × Tag2-Tracer stock solution, add to 1.98 mL 1×Detection buffer to dilute to 1×, mix and set aside.

4μL

Detection Reagent Mix

 

Take 50 μL 50× Eu-anti-Tag1 stock solution, add 2.45 mL 1×Detection buffer to dilute to 2.5 mL, mix;

Take 25 μL 100× SA-Ac stock solution, add 1×Detection buffer 2.475 mL to dilute to 2.5 mL, mix;

Mix both 1:1, this is Detection Mix.

10μL


 

1.2 Gradient Dilution of Test Samples

Taking VH032 as an example, the diluent is 1× Detection buffer. To reduce the interference of matrix effects, it is recommended to use a solution with the same matrix as the test sample for dilution; and adjust the test samples according to actual concentration.

Table 2. Positive Drug VH032 Gradient Dilution (Adjust According to Actual Situation)

 

VH032 Prepared Concentration (nM)

VH032 Final Concentration (nM)

Preparation Method

30,000.0

300,000.0

2μL 3mM Stock Solution + 18μL 1× Detection buffer

10000.0

100000.0

10μL ① +20μL 1× Detection buffer

3333.3

33333.3

10μL ② +20μL 1× Detection buffer

1111.1

11111.1

10μL ③ +20μL 1× Detection buffer

370.4

3703.7

10μL ④ +20μL 1× Detection buffer

123.5

1234.6

10μL ⑤ +20μL 1× Detection buffer

41.2

411.5

10μL ⑥ +20μL 1× Detection buffer

13.7

137.2

10μL ⑦ +20μL 1× Detection buffer

4.6

45.7

10μL ⑧ +20μL 1× Detection buffer

Blank

0.0

0

20μL 1× Detection buffer


 

 

2. Sample Addition and Controls

2.1 Sample Well (Sample): Add 2 μL test sample (gradient dilution), 4 μL Tag1-VCB protein working solution, 4 μL Tag2-Tracer working solution, 10 μL Detection Reagent Mix sequentially into the 384-well shallow well plate.

2.2 Maximum Signal Control: Add 2 μL Detection buffer, 4 μL Tag1-VCB protein working solution, 4 μL Tag2-Tracer working solution, 10 μL Detection Reagent Mix sequentially into the 384-well shallow well plate.

2.3 Negative Control (Negative control): Add 10 μL Detection buffer, 10 μL Detection Reagent Mix sequentially into the 384-well shallow well plate.

After adding all samples, centrifuge, seal the plate film, and incubate at room temperature for 2 hours.

 

Sample (Sample)

Maximum Signal Control

Negative Control (NC)

Step 1

2μL Test Sample

2μL 1×Detection buffer

10μL 1×Detection buffer

4μL Tag1-VCB protein Working Solution

Incubate at Room Temperature 10 min

Step 2

4μL Tag2-Tracer Working Solution

10μL Detection Reagent Mix

Seal plate with film, incubate at room temperature for 2h


 

 

3.Detection

Detect on a microplate reader compatible with TR-FRET. Excitation light is 320/340 nm, detect emission wavelengths of 620 nm and 665 nm.

[Result Calculation]

1) Calculate Signal Value (Ratio): Fluorescence signal at 665 nm divided by fluorescence signal at 620 nm, then multiplied by 10000.

Ratio = (665/620) ×10000

2) Calculate Net signal based on signal value:

Net signal = (Std-NC)/NC×100

3) Calculate CV (%):

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

 

[Data Example]

The data below cannot replace data obtained from experiments, it is for demonstration purposes only; results may vary depending on the plate reader instrument.

 

Note: Recommended Microplate (384-well plate, White, Shallow Well)

Shipping Notes
  • Free Standard Shipping on $100+ Orders to the USA.
  • Except Preorder products are shipped in 48 hours.
  • Delivery to the USA:
  1. Standard Shipping : 3-10 business days
  • If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
  • We offer a 30-day return/exchange service after receiving.
  • Final sale items are not eligible for returns or exchanges.
  • To process your return/exchange, please contact us at [email protected]
  • Please click here for more details>>> Return & Exchange Policy
SKU: 85937761780

Discover Niche Categories That Outsell

Top-Converting Item to Boost Your Average Order

4.8 ★★★★★
Based on 13 reviews
Sort
Highest Rating
Newest First
Oldest First
Product Reviews
J
Just Me
Los Angeles, US
★★★★★ 5
Room Divider That Gives Instant “This Is a Separate Space Now” Energy
Size: 3 Panel 12FT W
The Siebwin 3 Panel Folding Privacy Screen is surprisingly effective at changing the feel of a room without requiring construction, commitment, or somebody yelling about load bearing walls. The 12 foot width gives a substantial amount of coverage, making it useful for creating privacy, dividing open areas, blocking visual clutter, or setting up temporary sections within larger rooms. This saved us a ton of time and money in our new studio and enabled us to create a kitchen and office in one room. The wider feet make a huge difference because flimsy room dividers that collapse every time someone exhales too aggressively are deeply annoying. This one feels more stable and grounded once positioned. The folding design makes it easy to move around or adjust depending on the space, and it folds down relatively compactly when not in use. It is basically the introvert curtain wall of dreams. The material has a clean, neutral look that blends well into different room styles without screaming “temporary office cubicle panic.” It works well for apartments, studios, shared spaces, dressing areas, or simply hiding the mountain of chaos you swear you are going to organize tomorrow. Practical, portable, and refreshingly easy to use without turning setup into a three hour emotional event.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 16, 2026
S
Skeetre
Los Angeles, US
★★★★★ 4
It's 2 panels holding 1 big privacy screen. Can't use as 2 separate 4' panels, but works great.
Size: 2 Panel 8FT W
This 2 panel privacy screen works well. Assembly was very straight forward, with easily labeled tubes. My son and I had it together in about 10 minutes, which was probably 3 minutes of me telling him to quit playing around and hand me the parts. It feels very sturdy, not likely to fall over or anything. The material fits well, with the Velcro being very strong. Measurements are all very accurate, with all pieces attaching properly. We have a similar privacy screen at work that we have to try and stretch out because the top and bottom Velcro doesn't fit quite right... this model does not have any issues though. The material is thin and light weight, but solid black so that you can't see anything through or behind it. I definitely think it's a great deal for the money. What I think would have given it 5 stars though, is if you had 2 totally separate 4' panels instead of 2 panels with one big screen. It really wouldn't cost any extra to make it so that the screen was in 2 pieces and you could use it as 2 separate panels instead of having to use it as one big panel. It says 2 panel folding privacy screens, but it's really one big one that folds in the middle.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 25, 2026
B
Verified Purchase
Barbara Oleyar
Draper, US
★★★★★ 5
Beautiful cabinet
Color: White - 2 Doors
Love, love, love this cabinet!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 19, 2025
L
Verified Purchase
Leonard Wilson
Lexington, US
★★★★★ 4
Trouble hanging
Color: White - 2 Doors
This was easy to assemble, just the right size. The only thing was the way it was hung on wall, very hard to measure the right height.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 27, 2025
E
Verified Purchase
Elizabeth A Nelson
Grantham, US
★★★★★ 5
My 12 year old assembled this
Color: White - 2 Doors
This is a great organizational tool. We needed medicine storage in our bathroom and my 12 year old assembled it in about 20 minutes. Great instructions and plenty of shelving. It looks great in our bathroom. Just perfect easy storage
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 25, 2024

recommand products