Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 11 - Jul 16
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Human PHB 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 and centrifuge 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. Cell Supernatant: 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., 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 the 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. Preparation before testing: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the gradient working solution of the standard: Add 1 mL of universal diluent to the lyophilized standard. Let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. 3. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/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. 4. 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. 5. 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. 6. 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 uses 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 prohibitin (PHB) capture antibody. 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 prohibitin (PHB) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Prohibitin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Prohibitin, also known as PHB, is a protein encoded by the PHB gene. PHB genes have also been described in animals, fungi, plants, and unicellular eukaryotes. Prohibitins are divided into two classes, designated type I and type II, based on their similarity to yeast PHB1 and PHB2. Every organism possesses at least one copy of a single type of prohibitin gene. Prohibitins are evolutionarily conserved and ubiquitously expressed. The prohibitin gene, located on the BRCA1 chromosome at 17q21, was initially thought to be a negative regulator of cell proliferation and a tumor suppressor. This antiproliferative activity was later attributed to the 3' UTR of the PHB gene, rather than the actual protein. Mutations in PHB have been associated with sporadic breast cancer. However, overexpression of PHB is associated with decreased androgen receptor activity and reduced PSA gene expression, leading to reduced growth of androgen-dependent prostate cells. Levels of the longer transcript are higher in proliferating tissues and cells, suggesting that this longer 3' untranslated region may function as a trans-acting regulatory RNA. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, cell supernatant, tissue homogenate, etc. |
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
4.9 ★★★★★
Based on 694 reviews
Sort
Product Reviews
★★★★★ 4
Missing some ToC Pages
Format: Paperback
Item shipped missing the first couple pages from the Table of Contents. Otherwise, it is as expected. Using as AD Services reference in Enterprise IT office for junior admins to reference.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 21, 2026
★★★★★ 5
A great read for those who want in depth knowledge about MS Active Directory & related services
Format: Paperback
Reviewer: 20 yrs training and consulting experience with Windows Server, Exchange Server, Active Directory, all major hypervisors, and cloud services. Active Directory along with all the related services and features are complex and many faceted topics. Books showing step by step examples are easy to come by, although not very useful since there’s plenty of Microsoft online help with the same info. What’s more important is to understand the features and related services and how to use them properly and securely.
This book covers in great detail Active Directory and the related services and features. There’s also plenty of information and expertise from a security perspective. With the move to cloud services and more employees are working from home there’s a whole new level of security issues to contend with. This book contains lots of in-depth explanations that allow you to understand why certain features and services exist and how to use them. This extra depth and the author’s experience is what makes this book special.
I would recommend this book for IT Professionals that already have some AD knowledge and experience. My only qualification is that I occasionally found some of the explanations confusing even after reading them over a several times.
I would highly recommend this book for those who wish to understand, configure and design an AD infrastructure along with associated services including OU design, group policy, object management delegation, ADFS, Certificate Services, etc. Although the book is not about Azure AD it does cover some aspects including integrating with on-prem AD.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 10, 2022
★★★★★ 5
Detailed coverage of Active Directory
Format: Kindle
I spent several years very close to active directory but haven’t followed developments in recent years. I was interested in this book as a refresher on what’s new in the last 8-10 years – and it did not disappoint.
The chapter on Advanced AD Management with PowerShell does a great job providing some background, setting the stage for why we’d want to know this, and providing some brief descriptions of various cmdlets that might be needed. My last experience with PowerShell/Active Directory required the Quest module, so seeing these native cmdlets is a nice change.
The chapter on AD FS is very detailed, providing a history of the functionality, deployment topologies and considerations, as well as details on configuring the required components. This is a topic I was not familiar with before seeing this book, and it did a good job bringing me up to speed on the feature.
I have not made it all the way through this book, but the sections I have completed are very good. If you are looking for a thorough reference guide on the current state of Active Directory, you should check out this book.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 8, 2021
★★★★★ 5
The book for designing and deploying Active Directory in your Organization
Format: Paperback
If you purchased an earlier release of this book and was wondering if this one is worth getting, the answer is yes!! The biggest change is this edition thoroughly covers the latest and greatest, Windows Server 2022. It walks through security issues, the differences between this and previous versions of Windows Server and how to design and deploy a functional and secure Server 2022 Active Directory. I enjoyed reading through this book and highly recommend it!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 4, 2022
★★★★★ 5
Comprehensive guide for IT Professionals interested in increasing their Active Directory Knowledge
Format: Paperback
Dishan Francis is a seasoned IT expert and six-time Microsoft MVP who has offered innumerable lessons learned through his experiences using Microsoft services. The Third Edition of Mastering Active Directory provides simple explanations of ideas accompanied by appropriate graphics to help the reader feel at ease while learning new concepts.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 31, 2022