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Description
Human FITM1 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. |
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| 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 Fat Storage Inducing Transmembrane Protein 1 (FITM1). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Fat Storage Inducing Transmembrane Protein 1 (FITM1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Fat Storage Inducing Transmembrane Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Fat storage-induced transmembrane protein 1 (FITM1) is a protein encoded by the FITM1 gene. Recently, the fat storage-induced transmembrane protein 1 (FIT1/FITM1) family was discovered to be an evolutionarily conserved protein family involved in fat storage. In mammals, it is primarily expressed in skeletal muscle, raising the possibility that FIMT1 has unique functions. These proteins are specifically localized to the endoplasmic reticulum (ER) and, when overexpressed in cells or muscle, mediate the accumulation of triglyceride-rich LDs. Unlike triglyceride synthases of the ER-resident diacylglycerol O-acyltransferase family, FITs do not synthesize triglycerides but rather distribute them into LDs. | |||||||||||||||||||||||||||||||||
| 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. |
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| 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 |
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4.0 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 5
Excellent
Format: Paperback
While some readers may find John Paul II's writing style a bit circular or highly philosophical, the message presented in this text will change your life. Still, it is not easy reading. It is a collection of 129 audiences given by John Paul II commonly called the Theology of the Body. The audiences are based on Sacred Scripture. In this new translation audiences that were not delivered are now included. The Theology of the Body treats sexuality, marriage, celibacy, anthropology, family life, responsible parenthood, the wound of sin, redemption, and the resurrection of the body. This book is in my top five of best books.
I have read both translations and Michael Waldstein's is much easier to read than the first edition. The new headers at the beginning of each section that directly indicate what is being dealt with in the text. The introduction that Waldstein provides gives a theological and philosophical vision for understanding the text.
This book is worth your time. It stands against the culture of death and promotes the Gospel of Jesus Christ. It holds the answers the hunger that society is so desperately grasping at.
If you don't know much about the Theology of Body it would prove useful to read an introductory text by West, Hogan, or Percy & Schmitz. Christopher West, who is properly described as a catechist, has excellent presentations on the Theology of the Body in DVD and CD format that could prove helpful to a first time reader.
On a personal note: The Theology of the Body has helped me so much. I am in Holy Orders and I don't think I would be where I am except for this message (and God's grace and His will). This isn't some message that you can simply read and understand - it is something that you have to approach with an open heart and mind with prayer.
I really hope you read this book and promote it's message.
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Reviewed in the United States on September 28, 2009
★★★★★ 5
Stupendous and Life-Changing
Format: Paperback
I'll never forget the way I first heard the Theology of the Body. I was a brand-new sister, having made my first vows the year Pope John Paul II was elected. My community used to read a snippet or two from the Papal Audience talks right after breakfast, before we scattered to the four winds in carrying out the day's mission. Bit by bit, as we made our way through an unusual series of meditations on the book of Genesis, I found that the whole theme of "gift" was beginning to resound within me. It was like a new wine being poured into the wineskin of the Church.
Whatever our vocation, we are each called to be a "person-gift," as the Persons of the Trinity are "gift," given over unreservedly, and unreservedly received. In a vague way, even though I was still very young (!), I realized that something extraordinary was going to come from these talks, and that it would mean great things above all for women.
It took twenty years for that message to begin to seep into the language of our Church culture, but now small groups of young adults and couples are beginning to meet in homes and conferences to ponder John Paul's vast, fully Christian vision of humanity. I've used the Theology of the Body to present the essence of the vow of chastity to our novices and junior sisters, and I keep looking for ways to introduce it to others, especially to people who find themselves struggling to understand the Church's teaching in all those areas that touch on our vocation to love. The beauty of its truth is hard to resist.
This new and carefully researched translation offers more than just a consistent translation of key terms: previously unpublished units came to light; the Pope's original title and subtitles have been added; the whole content appears in the order in which the Pope meant for it to be covered. Yes, Dr. Waldstein's careful and scholarly introduction can be a bit overwhelming, but this book is worth taking the time simply to sit with.
That's what you do with beauty anyway.
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Reviewed in the United States on October 31, 2008
★★★★★ 5
Excellent Scripture-based compilation of lectures on human sexuality and marriage
Format: Paperback
This is a compilation of a series of lectures given by Pope John Paul II on the true meaning of sexuality and spousal union. It is a rather lengthy book, with lots of repetition because they are his lectures. At the beginning of each lecture, expect that he will "recap" what he discussed the previous time. It is good to read his actual words, but it requires patience to filter through the "new" material he presents because after a while you realize he is saying the same thing in different words; but, this may be required to get his message through to some readers/listeners. Each of us learns and grasps concepts differently, so his presentation of the material is geared toward universality in teaching the meaning of true spousal union as mutual self-gifting as given to us by God in the beginning before the fall in Eden. Using Scripture, JPII explains God's divine plan for human sexuality and its purpose; and he explains how objectification and selfishness in sexuality has led to a multitude of problems for society. He sees the sacramental aspect of marriage between man and woman through the eyes of Scripture. It is worth the time to read, but be patient with his repetitions, realizing that this was originally a lecture series. This book includes the "transcripts" of those lectures. Every Christian person--Catholic or not--should read his book to understand how sacred human sexuality is and how deviance destroys relationships.
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Reviewed in the United States on December 5, 2013
★★★★★ 5
A Catholic Must Read
Format: Paperback
Will change your complete view on life. Stick with it, it’s gold.
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Reviewed in the United States on March 3, 2026
★★★★★ 5
Great book
Format: Paperback
Great book
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 15, 2025