SKU: 28674002815

UniOne® TR-FRET Human BAFF/TACI Binding Kit

$328.50 $365.00
Save 10%
No.2 Bestseller on bieltoni.com >

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

Shipping Estimate
USA
  • USA
  • CAN

Ships within 48 hours · Estimated delivery Jul 17 - Jul 22

Promo Codes Available:

For Your Every Summer RSVP, with Code: SUMMER15

Description

UniOne® TR-FRET Human BAFF/TACI Binding KitProduct Specification Host Human Stability & Storage 80 Background The kit employs homogeneous time resolved fluorescence (TR FRET) technology to measure the interaction between Human BAFF and Human TACI. This method enables simple, rapid, and high throughput screening of inhibitors and antibody blockers. As shown in the figure, the interaction between BAFF and TACI is detected using an Eu labeled anti Tag1 antibody (TR FRET donor) and an acceptor

Product Specification


Host Human
Stability & Storage

-80℃

Background

The kit employs homogeneous time-resolved fluorescence (TR-FRET) technology to measure the interaction between Human BAFF and Human TACI. This method enables simple, rapid, and high-throughput screening of inhibitors and antibody blockers.

As shown in the figure, the interaction between BAFF and TACI is detected using an Eu-labeled anti-Tag1 antibody (TR-FRET donor) and an acceptor-labeled Tag2 antibody (TR-FRET acceptor). The binding of BAFF and TACI brings the donor and acceptor antibodies into close proximity, allowing the excitation of the donor antibody to trigger fluorescence resonance energy transfer (FRET) to the acceptor antibody, resulting in a specific 665 nm emission signal. The positive control drug Tabalumab blocks the binding of BAFF and TACI, preventing FRET signal generation. The stronger the inhibitory effect of the screened drug on BAFF and TACI interaction, the lower the signal. The signal intensity is proportional to the degree of BAFF-TACI interaction. No washing steps are required.

Components

组分

浓度

100T

500T

2500T

10000T

储存温度

Tag1-BAFF protein

100 ×

5μL

20μL

100μL

400μL

-80℃

Tag2- TACI protein

100 ×

5μL

20μL

100μL

400μL

-80℃

Tabalumab

25μM

5μL

15μL

75μL

300μL

-80℃

Eu-anti-Tag1

50 ×

10μL

50μL

250μL

1000μL

-80℃

Ac-anti-Tag2

12.5 ×

40μL

200μL

1mL

4mL

-80℃

Detection buffer

10 ×

400μL

2mL

10mL

40mL

-80℃

注意：所有首次解冻后立即分装，并按建议储存温度保存，避免稀释后储存及反复冻融。

Protocol

1. Reagent Preparation

1.1 Thaw all reagents at room temperature before use (allow at least 30 minutes for equilibration). The reaction system for the 384-well plate is 20μL (reagent volumes per reaction are shown in the table below). Calculate the required volume before preparation and prepare accordingly. 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

Take 2 mL of 10× Detection buffer and add 18 mL of deionized water to dilute to 1×. Mix well and set aside.

-

Tag1-BAFF protein

Take 20 μL of 100× Tag1-BAFF protein stock solution and add to 1.98 mL of 1× Detection buffer to dilute to 1×. Mix well and set aside.

4μL

Tag2-TACI protein

Take 20 μL of 100× Tag2-TACI protein stock solution and add to 1.98 mL of 1× Detection buffer to dilute to 1×. Mix well and set aside.

4μL

Detection Reagent Mix

 

Take 50 μL of 50× Eu-anti-Tag1 stock solution and add to 2.45 mL of 1× Detection buffer to dilute to 2.5 mL. Mix well.

Take 200 μL of 12.5× Ac-anti-Tag2 stock solution and add to 2.30 mL of 1× Detection buffer to dilute to 2.5 mL. Mix well.

Mix the two solutions at a 1:1 ratio to prepare the Detection Mix.

10μL


 

1.2 Gradient Dilution of Test Samples

Taking Tabalumab as an example, the dilution buffer is 1× Detection buffer. To minimize matrix interference effects, it is recommended to use a solution with the same matrix as the test sample for dilution. Adjustments should be made based on the actual concentration of the sample.

Table 2. Gradient Dilution of Positive Control Tabalumab (Adjust as Needed)

 

Tabalumab Preparation Concentration (nM)

Tabalumab Final Concentration (nM)

Preparation Method

①

2500

250

3μL 25μM stock solution + 27μL 1× Detection buffer
2. Sample Loading and Controls

2.1 Sample Well: Add 2μL of test sample (gradient dilution), 4μL Tag1-BAFF protein working solution, 4μL Tag2-TACI protein working solution, and 10μL detection reagent Mix sequentially into a 384-well shallow plate.

2.2 Maximum Signal Control: Add 2μL Detection buffer, 4μL Tag1-BAFF protein working solution, 4μL Tag2-TACI protein working solution, and 10μL detection reagent Mix sequentially into a 384-well shallow plate.

2.3 Negative Control: Add 10μL Detection buffer and 10μL detection reagent Mix sequentially into a 384-well shallow plate.

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

Sample

Maximum Signal Control

Negative Control (NC)

Step 1

2μL Test Sample

2μL 1×Detection buffer

10μL 1×Detection buffer

4μL Tag1-BAFF protein working solution

Incubate at room temperature for 10min

Step 2

4μL Tag2-TACI protein working solution

10μL Detection reagent Mix

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


3、Detection

Perform detection on a TR-FRET-compatible microplate reader. The excitation wavelength is 320/340 nm, and the 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 is for illustrative purposes only and cannot replace the data obtained in experiments. Results may vary depending on the plate reader. 

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: 28674002815

Discover Niche Categories That Outsell

Top-Converting Item to Boost Your Average Order

4.5 ★★★★★
Based on 12 reviews
Sort
Highest Rating
Newest First
Oldest First
Product Reviews
S
Verified Purchase
Searching the Stars
Grantham, US
★★★★★ 5
A Great Work on the Dynamics of the Human Condition
Format: Hardcover
Irvin Yalom, Professor Emeritus at Standford in psychiatry, has written a masterful book on the human condition (published in 1980). "Existential Psychotherapy" is singularly one of the best books I have ever read. Why? Yalom has combined 1) his intellectual insights in psychology with 2) his practical observations in therapy with 3) existential thought. He seeks to show that much of the anxiety we face as human beings, and seek comfort for in psychotherapy (or religion), originates from the existential facts of being a mortal, free being. He focuses on four main areas: death (our inner structure will go to nothing), freedom (we are forced to make momentous decisions without full information), isolation (we are alone in many fundamental ways) and meaninglessness (we don't have a clear picture of what it really means). From literature, psychology, philosophy and some theology (Tillich), Yalom has created a definitive text on what it means to open our eyes fully to our existential situation. It is not a depressive text, as the goal is to come to terms with our condition. And ironically, facing the truth of death, for example, makes life richer. This is much more than a self-help book, it is a philosophical-psychological masterpiece. I have a Ph.D. in the philosophy of religion and have published several articles in leading journals and don't give out the kind of praise I am giving here easily. It is one of the best books I have ever read, and I think a wide range of people would profit from it. Yet it's a difficult book, not an easy self-help read. But Yalom is such a clear writer that most readers could follow it. When he uses technical psychological or philosophical terms, he usually explains them. Otherwise, the dictionary must be used. This text would be helpful for people in the helping professions in giving an existential aspect to their work. Religious thinkers and those interested in philosophy should pick it up at any cost. Any educated person could profit from it greatly. It's a long text, and I had to break it up by carefully reading some from each of the main four parts simultaneously to get a view of the whole. It's a bit dry at times, but thoughtful, perceptive, educational in many fields, --it's ....wisdom literature....I can't think of a higher compliment.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 19, 2013
D
Verified Purchase
David L. Pogge, Ph.D.
Chelsea, US
★★★★★ 5
The best of its kind.
As a clinical psychologist who has been practicing, doing research on, and teaching assessment and therapy for more than 25 years I have encountered many books and many more articles and research papers on psychotherapy. While some have real value, most do little more than take up space and help the people who wrote them advance their careers. All too rarely one comes across a book that changes one's way of looking at things and helps one to approach this field in a new and qualitatively better fashion. Yalom's book on Existential Psychotherapy is one such book. I first read this when I was in graduate school in the early 1980's and just learning about psychotherapy, and about life. This book opened my eyes and gave me an over-arching way of thinking about myself, my patients, and the things that I was doing. Unlike most texts on therapy, it was not limited to the parochial "theoretical orientations" and mechanistic models of therapy technique that I was learning then and have seen proliferate ever since. Since this time I have re-read this book many times, and each time I have gotten more from it. Yalom tries to go through the collected wisdom of mankind, not just in the narrow world of scientific psychology, but in philosophy, art, and religion, to explain how everyone must face certain existential realities such a mortality, temporality, resonsibility and isolation, and how the struggle to face these basic issues underlies many of the symptoms and complaints that lead people to seek the help of mental health professionals. Unfortunately, as Yalom points out, all too often we fail to see these as such, and offer help in the form of superficial, pseudoscientific psychologizing, technical trivialities and medications that often do more harm than good. While, used humbly and wisely, these things are not without value, their value is often fatally limited by their failure to see the larger context of life and its challenges. Once having read and really thought about Yalom's book, that becomes much harder to do; and thereby inevitably enriches one's practice and one's life. Regardless what kinds of therapies and what kinds of patients one works with - or even if one's practice is limited primarily to assessment or research - this book provides a philosophical and conceptual bedrock that can inform and enrich what one does. I am so happy to see that this book is still in print, and I would encourage anyone who plans to make a living trying to be seriously involved in other people's lives to avail themselves of this book. It is a treasure and a masterpiece. I intend to us it in my classes this year and for as long as it remains in print.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 5, 2013
C
Verified Purchase
Classical Fan
Lexington, US
★★★★★ 5
Very Interesting and Well-Written
Format: Kindle
This is a very interesting and well-written book that discusses philosophy as well as psychology, while also providing numerous vignettes from actual cases, that demonstrate the implementation of the author's ideas. Highly recommended.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 9, 2024
P
Verified Purchase
Pat
Battle Creek, US
★★★★★ 5
This is a really great book.
Format: Hardcover
I am a layperson so those who read this review should consider that. I have read this book slowly, very slowly and as a layperson have found it to be so very, very enlightening. This material has such depth and there were sections that I read that caused me to put the book down and consider what I had just encountered. Life is not a simple journey especially when we pick up so much detritus on the way. For years, I knocked on so many variously colored doors - entering and finding myself wanting...not them, mind you, but myself. Reading this - pointed me to a gate that swung open to a garden area, albeit, very overgrown, untended. It is here now that I work - some of what I thought were weeds are not. Some of what I thought were flowers are not. I just want to be free and am willing to accept that responsibility. Thank you, Dr. Yalom....you sage of my generation.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 7, 2014
D
Verified Purchase
drey c.
Bozeman, US
★★★★★ 4
Great book for growth
Format: Hardcover
Deep and thoughtful book, worth the investment for any training psychotherapist or social worker. Yalom is a leader in the field and one whom I trust. I have my PhD in psychology and can't suggest this enough. .
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 7, 2015

recommand products

Cisco SFP-10G-ER-S 10GB BASE-ER Extended Reach SMF Optical SFP+ Transceiver
139.99
Pretty Cure Kimi to Idol Special Good Job Sticker Set
11.92
Rilakkuma Face Plushie - Blue Wolf / Mini
14.90
Sanrio Yokai Light-Up Pen - Hello Kitty Nekomata
14.90
Rilakkuma's Lunchtime Bento Box
27.90