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Description
Human ZFAND6 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 Zinc Finger, AN1-Type Domain Protein 6 (ZFAND6). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Zinc Finger, AN1-Type Domain Protein 6 (ZFAND6) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Zinc Finger, AN1-Type Domain Protein 6 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | AN1-type zinc finger domain protein 6, also known as ZFAND6, is encoded by the ZFAND6 gene. It is involved in regulating TNF-α-induced NF-kappa-B activation and apoptosis. It also regulates the Lys-48-linked polyubiquitination status of TRAF2 and reduces its association with RIPK1. It is required for PTS1 target sequence-dependent protein import into peroxisomes and for PEX5 stability, and cooperates with PEX6. In vitro, it participates in the export of PEX5 from the cytosol to the peroxisome. Diseases associated with it include peroxisome biogenesis disorder 1B and Zellweger syndrome. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.1 ★★★★★
Based on 1806 reviews
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Product Reviews
★★★★★ 5
Crossbody Purse with a choice of a bunch of colors to choose from!!
Color: Red
This is a great crossbody purse for anyone!! It holds my wallet, makeup bag, phone as well as a ton more space to add more stuff in!! I currently own the red and the army green. This is a great purchase!
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Reviewed in the United States on November 23, 2025
★★★★★ 4
Decent bag
Color: Light Grey, Color: Light Grey
2.5 year update: I'm ready to replace my beloved grey bag, but most of the colors more than doubled in price!! I get that there's inflation, but that's insanity. It was $10.99 when I bought in 2019. Forget this brand. Dropped a star.
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Original review:
In the photo, that's a 32oz Nalgene bottle and a small, Periea Chelsy organizer (found on Amazon, 8.5 inches long). 9 months of near daily use so far, no issues! I even got oil and ink stains on it, and they came right out.
Great runaround bag, not too big not too small. Love it. First things I did, removed the label and coated it in Kiwi Protect All spray.
Pros:
- 2 pockets inside are deep and sooo useful
- very cute
- comfortable and very light to carry
- adjustable strap length, for shoulder or crossbody
- price is right
- machine washable
Cons:
- zipper is a bit "sticky"
- words on the label are classic Chinese-English, reminds me of Taiwan. Lol
- no inside lining
- fabric could be thicker. It's thin for canvas.
- metal hardware is not stainless steel
Neutral:
- medium size (model must be child size, pay attention to dimensions. I'd say it's similar to standard disposable grocery bag size, cept taller)
- durability is questionable. It's not going to come apart if you look at it wrong, but I definitely wouldn't try using this as a bookbag either. For the price, it seems sturdy enough.
When this bag dies, I will 100% be buying another one if it's still available. Love it.
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Reviewed in the United States on February 15, 2019
★★★★★ 5
Great value for amazing bag
Color: Pink
Great deal on this tote bag, the strap length is extremely adjustable for size and different wearability. It has good storage space and no pockets inside but works well. It looks great with different colors I have the pink and my boyfriend has the coffee one which looks amazing. I added pins to mine and looks great! Love this bag
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Reviewed in the United States on October 10, 2025
★★★★★ 3
I love my bag.
Color: Leopard White, Color: Leopard White
love the design, i barely put anything in it tho because it weighs the bag down even though it’s a nice size. out of the package looks like it hasn’t been sown properly thru, pieces of the fabric sticking out. (loose thread) you get what u pay for but still pretty.
Update, Maybe i underestimated my baby love, she fighting the greatest fight! Still love my baby! I would buy another! i want the whole collectionnnnn!
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Reviewed in the United States on September 12, 2025
★★★★★ 5
LOVE THIS ZEBRA BAG
Color: Zebra White
I absolutely LOVE this pocketbook! I searched for a zebra print bag, then finally found this! It's big, so beware... Now l lug around everything, haha! Ive had it around a year now & it's traveled alot! It has held up well, except for a few threads, but lve seriously got wayy too much weight in mine! I do wish it had more than the little pockets on the inside (Suggestions for future bags! A big side, zippered pocked would be nice) But lm so glad l got it!!
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Reviewed in the United States on December 1, 2025