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Description
Human BACH1 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend them 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, the PBS volume can be reduced appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cerebrospinal Fluid Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 transcription regulator protein BACH1 (BACH1). 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 transcription regulator protein BACH1 (BACH1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Transcription regulator protein BACH1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The transcriptional regulator BACH1 is a protein encoded by the BACH1 gene. This gene encodes a cap-n-collar type transcription factor belonging to the basic region leucine zipper family (CNC-bZip). The encoded protein contains extensive complex, triad, triad/poison, and zinc finger (BTB/POZ) domains, which are atypical among CNC-bZip family members. These BTB/POZ domains facilitate protein-protein interactions and the formation of homo- and/or hetero-multimers. The C-terminus of this protein contains a leucine zipper from the bzip_maf family. When heterodimerized with MafK, this protein functions to inhibit Maf recognition elements (MAREs), resulting in transcriptional repression. Multiple alternatively spliced transcript variants have been identified for this gene. Pathways implicated in this gene include the comprehensive breast cancer pathway and the BRCA1 pathway. Gene ontology annotations associated with this gene include DNA-binding transcription factor activity and heme binding. An important homologue of this gene is BACH2. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cerebrospinal fluid, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 5
A readable set of reflections on Faith, Hope, and Charity
Format: Hardcover
Cardinal Raniero Cantalamessa offers a series of wonderful theological and spiritual insights into the three theological virtues of Faith, Hope, and Charity. Taking presentations on each of the virtues, the Cardinal edited each into smaller units, each three to four pages in length. This makes for easy reading (one could take a selection a day) for "lectio divina". His treatment of "Justification by Faith" puts what has been a "thorn of contention" into easily understood terms that can open itself to ecumenical dialog with other Christian denominations for whom this has been a point of misunderstanding of the Catholic position.
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Reviewed in the United States on October 18, 2024
★★★★★ 5
We can all be wise men bearing gifts to Jesus
Format: Hardcover
Faith, hope, and charity are not just virtues that we can develop to bring ourselves closer to God and our fellow person but they are also gifts that we can bring to God, writes this household preacher to two Popes. Cardinal Cantalamessa writes on serious topics with an accessible and joyful style that welcomes the reader to see him or herself as one of the Magi bringing precious and deeply meaningful gifts to Christ.
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Reviewed in the United States on November 14, 2024
★★★★★ 5
Cardinal Cantalamessa is a Saint!
Format: Hardcover
This book has the wisdom of the fathers infused with the gentle Grace of the Holy Spirit, written for modern day Christian readers.
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Reviewed in the United States on December 20, 2024
★★★★★ 4
Beautiful and profound, albeit flawed
Format: Hardcover
What I'd really like to rate this is 4 1/2 stars, because it isn't perfect, but it's better than a 4. I just can't bear to give it a 5, for reasons described below.
The book has three main parts, each named for a theological virtue of the title. Altogether there are 40 chapters: 14 dedicated to faith, 10 to hope, and 16 to charity. A couple of "Bonus" chapters called "Excursus" take up some interesting theological questions that are related but don't quite fall under any one topic: did Jesus possess the theological virtues? and From God as Love to the Filioque.
The book is deeply learned and cites theologians through the centuries, including a few I'd never heard of despite a lot of formal and informal theological study. Of course you meet the usual suspects such as Origen, pseudo-Dionysius, Augustine, Thomas Aquinas, Catherine of Siena, John of the Cross, Mother Teresa, and others; but: have YOU heard of St. Zeno of Verona before? If not, you're in for a treat!
The text has more of an Augustinian flavor than a Scholastic one. (I should qualify that claim by admitting that I am at best an armchair theologian, so don't quote me on that.) But, for example: Chapter 39 discusses Beauty, uses the idea of God's eternal Beauty to explain the Trinity, and concludes with St. Augustine's "recipe" for becoming beautiful. Indeed, an in-depth discussion of the Trinity doesn't appear at all until Part 3 on Charity, rather than in the chapter on Faith!
An important distinction that Fr. Cantalamessa draws is between "objective" aspects of a theological virtue and the "subjective" aspects of the same. The "objective" aspects refer to the object of the virtue, such as God Himself; the "subjective" aspects refer to how we experience them. He points out that, historically, the "objective" view tended to predominate in theological discussion, whereas the "subjective" view tends to dominate more recently, almost to the exclusion of the objective. It will probably not surprise the reader that Fr. Cantalamessa, former Preacher to the Papal Hosuehold, comes down squarely in the Catholic approach of "both/and", and he elaborates on this.
A very appealing aspect of this book is the occasional use of parable and analogy to explain difficult subjects: a parable on trying to justify ourselves by our works (Ch. 7), analogies for "the hint that God exists" (ch. 5), the analogy of the seed (ch. 14), an analogy that hope needs difficulties and tribulations (ch. 22), the Trinity (ch. 29), God's love for us is erotic (ch. 34), and how we might hope that even Nietzsche can be saved (ch. 37). Many analogies, though not all, are drawn from ordinary family life: a mother's love for a child, a child's temper tantrum before collapsing in tears on a parent. These are powerful and effective.
Unfortunately, I can't rate it 5 stars, because the text seems to consider its audience to be the average educated lay Catholic, but there are two serious weaknesses both for the theological newbie and even for the theological adolescent.
One is the use of many unfamiliar terms, some of them merely transliterated from Greek, and no definition given anywhere -- often, not even a hint of what the word may mean. In some cases this can make it difficult to follow the discussion. For instance, the text dedicates two entire chapters to the question of justification, which makes sense given that it's an important topic in the realm of faith, and it's important to take it seriously. But the book never once provides a definition, which suggests the reader should be familiar with the term already. I guarantee you most people don't know what the word means. But even if you think that a definition of "justification" will indeed come tripping off the average reader's tongue, I challenge you to make a case for terms like ontological, parousia, and parenesis. I've been reading Catholic theology for 30 years and parenesis is a new one even to me. Sure, the reader might could look them up, and I'm glad to expand my vocabulary, but who's the audience here? If the text is meant only for seminarians, then never mind, but given how Word on Fire is marketing this I really don't think that is the case.
The second major weakness is all the more disappointing, as it is so common to contemporary works of theology: when newer developments seem to contradict past dogma or even Scripture itself, pretend the dogma and Scripture doesn't exist. This happens at least twice:
1) Surely Fr. Cantalamessa is not unaware that Scripture both Old and New is replete with references to Christ "ransoming" us from God's wrath. Yet there he is in Chapter 31, not merely acting as if it doesn't exist, but contemptuously dismissive of the notion! Grant the Scholastics this much: at least they took Matthew 20.28, Romans 1.18, Romans 2.5-8, 1 Timothy 2.6, 1 Peter 1.18-19, and Revelation 19.15 seriously enough to wrestle with them.
2) Similarly, the Council of Florence made certain pronouncements on "those existing outside the Catholic Church" and "the souls of those who depart this life in actual mortal sin, or in original sin alone." Chapter 14 acts as if they do not exist. I do not for a moment mean to advocate for the "glass half-empty" interpretation of these pronouncements that predominated theological discourse for centuries, let alone for Feeneyism, but we ignore them at our peril, if only because ignoring them leaves a great big breach in the apologetic wall that will come under assault both from those who do reject the Second Vatican Council and from hostile Protestants more knowledgeable of Catholic theological history than the average Catholic and, one half-wonders, the average Catholic theologian.
Those drawbacks, while severe enough in my eyes to warrant mention and deduct a star, do not for a moment take away from the beauty and profundity of the rest of this work. I am very glad to have had the chance to read it; it has challenged me both intellectually and spiritually, and I have given it to my (late teenage) children to read and discuss with me. It is absolutely worth reading, and you WILL get a lot out of it. Just be ready for the challenge.
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Reviewed in the United States on July 25, 2025
★★★★★ 5
OPEN YOUR EYES
Format: Paperback
Great book! Open your eyes sheep..... there is much more to this world that we know of. Authority figures have lied to us long enough..... do your own research. This book is a great example of what we were not taught in school! Well written.
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Reviewed in the United States on February 14, 2021