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Description
Human HDAC6 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 freeze-thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes and remove the supernatant for analysis. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Preparation for Assay: 1. Remove the reagent kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. To 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 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 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 histone deacetylase 6 (HDAC6) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of histone deacetylase 6 (HDAC6) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Histone Deacetylase 6 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Histone deacetylase 6 is an enzyme encoded by the HDAC6 gene. Disorders associated with HDAC6 include chondrodysplasia with lamellar processes, distinctive humeral processes, hydrocephalus, and microphthalmia and humeral processes. Pathways associated with HDAC6 include activated PKN1, which stimulates transcription of the AR (androgen receptor)-regulated genes KLK2 and KLK3, and the BRCA1 pathway. Gene ontology annotations associated with this gene include enzyme binding and ubiquitin-protein ligase binding. An important homolog of this gene is HDAC10. | |||||||||||||||||||||||||||||||||
| 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 homogenate, cell culture supernatant |
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4.4 ★★★★★
Based on 750 reviews
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Product Reviews
★★★★★ 5
A superb addition to the Marvel Darth Vader canon
Format: Paperback
An almost perfect run of comic books, focused around the Imperial assault on Mon Cala post-Episode III. Many familiar faces are here: Vader, of course; Tarkin; Ackbar; Palpatine; Rogue One’s Admiral Raddus; and many more, including Sith, Jedi, and clones (there’s a nice nod to Order 66). Charles Soule deftly weaves an intriguing narrative, fast-paced but also with plenty of depth. The art is also excellent. In addition to the main stretch of the story, the trade also contains the Darth Vader annual, an outstanding Rogue One story that caps things off nicely. Highly recommended reading for Star Wars fans.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 17, 2019
★★★★★ 4
Good for Darth vader fans.
Format: Paperback
This book was amazing. I prefribly like vol.4 more than vol.3. The writers of this book have outdone themselves again, another one of Vaders archaic stories and the book was in perfect condition. I highly recommend it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 10, 2025
★★★★★ 5
Charles Soule's Series Continues to Impress and is Arguably the Best of Marvel's New Canon
Format: Paperback
Darth Vader leads the invasion of Mon Cala. With the Empire tightening its grip across the galaxy, Vader is dispatched by his master to the aquatic world of Mon Cala to track down a rogue Jedi who may be advising the planet's king. With the Inquisitors in tow, Vader and Grand Moff Tarkin face off with one of the first open acts of rebellion in their new Empire. Soule is at his absolute best with this series as he continues to explore a version of Vader haunted by his own inner goodness and memories of the past (the book does contain some references to events of the Clone Wars television show, but you don't need to have seen it to grasp the story). Likewise, his exploration of the seduction of the dark side is fantastic with a new Jedi target willing to use deception and war in order to light the sparks of a future rebellion. The final issue of the book may be one of the best Star Wars comics of the entire new canon with Tarkin hunting Vader for sport (on the latter's request oddly enough) across a desolate and hostile planet. The issue isn't what one expects but makes a great deal of sense when exploring the relationship between these two characters (while also explaining why Vader is so deferential to Tarkin during A New Hope). The final addition is a decent annual that sees Vader investigating the Death Star and sabotage of its construction. The annual isn't the best addition to the series (the artwork isn't up to the standards of the rest of the series), but it does introduce some intriguing ideas about Tarkin and Vader's relationship and the events that set up the Rogue One movie.
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Reviewed in the United States on September 17, 2018
★★★★★ 5
Nice
Format: Kindle
Love reading comics on the Kindle this is a great story leading up to the creation of the first death Star
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Reviewed in the United States on October 31, 2024
★★★★★ 5
Good read
Format: Kindle
Lots of questions answered. Worked in the canon beautifully. Suggested read for every Star Wars fan. I highly recommend it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 29, 2023
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