Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Aug 7 - Aug 12
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Human PIAS3 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre chilled PBS (0. 01M, pH 7. 4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and
Product Specification
| Usage | Required experimental equipment:
1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
|||||||||||||||||||||||||||||||||
| Theory | This kit utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against the E3 SUMO-protein ligase PIAS3 (PIAS3). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of E3 SUMO-protein ligase PIAS3 (PIAS3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human E3 SUMO-protein ligase PIAS3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Protein inhibitor of activated STATs 3 (PIAS3), also known as the E3 SUMO-protein ligase PIAS3, is an enzyme encoded by the PIAS3 gene. PIAS3 was initially discovered to interact specifically with phosphorylated STAT3 in interleukin-6 (IL-6)-activated myeloblast M1 cells. This interaction is mediated by PIAS3 binding to the STAT3 DNA-binding domain. Consequently, STAT3 transcriptional activity is inhibited by physically preventing its binding to target genes. Subsequently, it was also discovered to be a regulatory protein of other key transcription factors, including MITF, NFκB, SMADs, and the estrogen receptor. It also functions as a SUMO (small ubiquitin-like modifier)-E3 ligase, catalyzing the covalent attachment of SUMO proteins to specific target substrates. It directly binds to several transcription factors and either blocks or enhances their activity. Alternatively spliced transcript variants of this gene have been identified, but the full-length nature of some of these variants has not yet been determined. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
|||||||||||||||||||||||||||||||||
| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- 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
4.0 ★★★★★
Based on 16 reviews
Sort
Product Reviews
★★★★★ 5
The most impactful book on lasting love & relationships
Profound - highly recommend this book to EVERYBODY who has or wants to have a significant other. The different love languages really resonates with me and could save many relationships. This book initially a gift to me from my childhood best friend. It practically saved her marriage. I have since read it and purchased it for other loved ones.
Dr. Chapman explains how important it is for couples to understand how each other and themselves both give and receive love. It is possible for couples to truly love each other, but to truly feel unloved because they don’t think the same about giving and receiving love. Everybody generally has their own primary love languages for receiving love and giving love. It may be the same for giving/receiving, and it may be different. If a husband does not meet the primary love language of his wife, she might not sense his true feelings and start to be unsatisfied with their relationship.
Understanding your spouse’s love language and acting accordingly will fill their “Love Tank”. The “Love Tank” analogy is a great metaphor for describing how loved someone feels. Meeting people’s primary love language consistently will fill up their love tank and help them feel loved like they need. But if a spouse fails to meet this primary love language, it might leave their “Love Tank” empty, which leads to feelings of being unloved and issues in relationships. Secondary languages are also important, so it's critical to reflect and understand your own priorities and that of your spouses.
The five love languages are:
1. Words of Affirmation: If this is your love language, you feel most cared for when your partner is open and expressive in telling you how wonderful they think you are, how much they appreciate you, etc.
2. Acts of Service: If your partner offering to watch the kids so you can go to the gym (or relieving you of some other task) gets your heart going, then this is your love language.
3. Affection: This love language is just as it sounds. A warm hug, a kiss, touch, and sexual intimacy make you feel most loved when this is your love language.
4. Quality Time: This love language is about being together, fully present and engaged in the activity at hand, no matter how trivial.
5. Gifts: Your partner taking the time to give you a gift can make you feel appreciated.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 7, 2014
★★★★★ 5
Helped my marriage
First I must say I REALLY enjoyed this book. Me and my husband both. I heard about this book on Moody radio and decided to give it a try because I wanted a better marriage with less fighting and disagreements. I am soo happy I did. This book was right on point for me and my husband. I read it first, then got the audio version for him to listen to, which he did 4 times. It really gave me great insight into my self and my husband. It helped me to understand how to best express my love to my husband in the way he wants and understands most, and he was able to do the same for me. This hasn't been a cure all, but it helps to understand each other much more than we ever have before and we have been together for 18 years.
The book is very well written, its an easy read and you should are able to get through it quickly. The change comes with investing time to apply the principles you have learned. I personally had to go through the material more than once to really let it sink it. This has been a small financial but HUGE emotional investment in one of my most important relationships. The knowledge in this book has really help my husband (his words) to better navigate personal relationships, not just with me, as it is intended, but also with his sister, friends and even stranger. I have found I can use this information is so many interactions and encounters with people throughout the day, it really opens you up to a new perspective. Gary Chapman did a great job explaining the details of the love languages. Anytime in conversation with someone complaining of relationship or even just communication issues I make sure to recommend this book. Can't say enough good things about it and we plan on checking out his other books as well.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 12, 2016
★★★★★ 5
This saved my marriage
Really, it did. Simple and profound, a quick read. We all want to express love in the form that we want it for ourselves which is a recipe for disaster and completely arbitrary for your partner however well meaning that is. My wife kept saying that she didn't feel loved in spite of my significant attempts.... Now I know why. For instance saying "I love you" had absolutely no impact on her because her "words of affirmation" category is zero (absent). But she has a high need for physical touch (hooray for me because that is a big match). That insight lead to further research into tantric sex and now I'm having the best sex of my life and more frequently than when we were younger (amazing on both counts). The key was finally understanding what she needed so that she could feel "filled up" In the customized way that she needed.
The examples are a bit dated, but still conceptually valid. For her the "acts of service" wasn't washing the dishes, but acts of targeted thoughtfulness that took some time to properly distinguish. I took it on to do something appropriately thoughtful for her every single day since she tested high in that category.... That was a grand slam home run over time.
We also took a course in the enneagram (highly insightful personality typing) about the same time that dovetailed nicely. She was a type 2 that wants to make everyone around her happy, everyone except herself of course; she gives and gives until she is depleted and then becomes resentful. For her to be able to state what she wants and needs remains a huge struggle for her but she expects me to just know... A paradox for sure, but now I understand that by keeping her "filled up" overcomes that sense of depletion. (The enneagram is also highly recommended to know yourself and those around you). She takes care of those round her and she needed someone to do that for her; a huge insight.
The punch line is that I now get back what I need with a new passion that feels more like an ongoing honeymoon. Priceless. ;-)
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 11, 2014
★★★★★ 4
As of July 2012 - 92% 4 & 5 star reviews
As of the time I am writing this review 368 out of 398 reviewers gave this book a 4 or 5 star rating - that's 92% "I liked it" and "I loved it" ratings. With these many positive reviews there are some critical reviews as well that are worth reading to get a balanced overall review - there may actually be more (and likely are more) than 5 love languages or categories. The author has a significant amount of knowledge and experience regarding married couples and it is certainly worth considering his input. What will make the information in this book the most beneficial is incorporating it with personal experience, and this subject will likely be a "work in progress" project with a focus on getting better everyday to result in a lasting, happy, and fulfilling marital arrangement.
My favorite review is "Learning to Speak, December 23, 2010" where the reviewer's review could have been a superb foreword for this book. May I suggest reading it as in my opinion it is brief, clear, and simple. If you have time consider reading the other reviews and comments too.
Of course, some may not agree or totally agree with this book's author; however, the subject of marriage is simple, yet complexed - and even compounding at times.
In my opinion this is one of the better books on this subject. There is some good material here making it worth considering reading it.
This book did stimulate my thinking on the different viewpoints in marriage and if you'd like to read my comments on this marriage subject contine, if not please feel free to move on. I am just hoping that some of these thoughts may help some considering marriage or who are already married.
Some believe that men and women basically use different parts of their brains. Often heard are: "The left brain thinks, the right brain feels." "The left brain analyzes, the right brain intuits." "The left brain is logical, the right brain is emotional." Likely, our thinking, feeling, and loving are more complex than these simple statements; yet, at least on occasion (likely more often) men and women think and feel differently and express themselves differently - the author of this book identifies, categorizes, and classifies love into five languages. I would add one additional language, which is the ability to sincerely and promptly say "I'm sorry" from one's heart. From my 45+ years of marriage and from what I have learned from many others, a successful, lasting, and happy marriage involves two great forgivers and apologizers. In my three and a half decades of managing people I have found that those who never or almost never say "I'm sorry" have difficulties with their working and personal relationships.
A husband and a wife differ to varying degrees about how they both think and feel about things, and this is in harmony with how the Creator said regarding Adam that He was going to make a helper for him, as a complement of him (not an identical twin of him - she was made different in a good way). A complement completes, perhaps making something just right. A husband and wife will benefit from loving each other, especially as the other person wants and needs to be loved.
Couple this with deep respect and you hold the two keys to a successful, lasting, and happy marriage and family life - Love and Respect.
Hopefully adding this thought will help your loving and respectful marriage grow more each and every day: "I love you more today than yesterday, but only half as much as tomorrow."
And one additional thought: "It is more beneficial for me to be respectful and loving in all that I do, than for me to be loved (something I very much want)."
Every marriage has the potential to be successful, lasting, and happy, especially using the two keys of "Love" and "Respect." Your marriage can be a most precious, valuable, and wonderful gift by using these two keys with sincerity and heartfelt caring; and, never let pride, the childish silent treatment, or other unloving disrespectful traits mar your treasured marriage!
A good "PRIDE" antidote expressed before the end of the day: "I'm sorry - I was mistaken - How can I make it up to you? - I'll do my best to be better - Will you please forgive me?"
A good "CHILDISH SILENT TREATMENT" antidote as soon as possible: Rescue the loving, caring, and respect adult within you. "Whining" and "I won't talk to you" are childish - they rarely worked in childhood and have no place among true adults. "Scolding" and "Lecturing" is easily blocked out. The best communications are loving, caring, and respectful adult expressions coupled with a big dose of attentive listening and understanding.
In ballroom dancing it has been said that "it takes two to tango," and "it takes one to lead." Many have found a successful, permanent, and happy marriage includes three - the loving husband, the respectful wife, and the Creator and Author of marriage (who perfectly knows what's best).
A good question to ask yourself at the beginning of each day: "What will I do today that shows I both love and respect my spouse?" TIP: While certainly one positive act or action daily is a good start, many are even better and will bring more benefits.
ADDITIONAL BENEFICIAL READING: "One Minute for Myself [Yourself]: How to Manage Your Most Valuable Asset" by Spencer Johnson, MD - while it is good to have a great relationship with your spouse; it is essential to have a good relationship with yourself, especially if your goal is to love your neighbor as yourself. Keep in mind if this is one of your goals that your closest neighbor is your spouse. Good relationships with ourselves and others I believe is what our true success in life is all about. My thought is that one needs a good relationship with oneself first in order to have good relationships with others - and it is wise to pursue "self-respect" by being respectful of yourself and all others. I like the thought of "self-respect" rather than "self-esteem" because it is easily possible to think too much of oneself; better to just focus on being respectful, caring, loving, and having proper self-respect.
ADDENDUM: One of best ways to tell your spouse "I Love You" is to say "I love you just the way you are." The principle here is if you want to be accepted in any relationship you should give your acceptance first. How many of us really want someone to relentlessly badger us to change this or change that about ourselves. Change in itself can be difficult, but that is another subject to consider.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 11, 2012
★★★★★ 5
A Way to Divorce Proof Your Marriage
"The object of love is not getting something you want, but doing something for the well-being of the one you love." Statements like this and many others are the treasures that fill this book. This book is a must read for anyone who is married or even considering it. It is full of real life accounts from people who had problems in their marriage, but eventually overcame them. These stories give not only ideas on what to do, but inspiration and desire to build a strong marriage. The 5 Love Languages are something Gary Chapman came up with after years of marriage counseling. He didn't come up with these out of thin air, he had so many experiences with relationships and discovered common love patterns among spouses. He concludes that there are 5 different languages of love that people speak. A love language is the way a person feels love from another. That could be through acts of service, or physical touch. Discovering the way your spouse feels love will save a relationship. I thought to myself, "Ok, the 5 love languages are listed on the back cover; what's the point of reading it now?" But after reading in depth about each love language my eyes have been opened on exactly what I must do to accommodate my wife's love language. The book gives so many examples; at least one of them is sure to be your case. If you didn't realize what you were doing wrong, the examples will spark that within you. I took notes and underlined many passages. At the end of each chapter he asks an open ended question to make you think about how you can apply what was discussed. This book is the service manual for any marriage. Study and apply what you read and I can assure you a full "love tank" leading to a better marriage.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 13, 2013
recommand products
Psy 4 de la Rime: 4Eme Dimension (Digipack LTD) - COMPACT DISCS
26.99
Sir Path tik: 100 000 Fois Merci - COMPACT DISCS
19.99
Mgm Rockabilly: MGM Rockabilly - COMPACT DISCS
27.99
Luc De Larochellière: C Est D L Amour Ou C Est Comme - COMPACT DISCS
20.99
Morning Episodes: Never Felt So Alive - COMPACT DISCS
27.99