SKU: 64466293908

Mouse Col6a1 ELISA Kit

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Description

Mouse Col6a1 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 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 freeze-thaw cycles 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 1000×g for 20 minutes, and collect the supernatant for analysis.

Preparation for the Assay:
1. Remove the reagent 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 stand for 15 minutes to completely dissolve, then gently mix (concentration 40 ng/mL). Then dilute to the following concentrations: 40 ng/mL, 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 40 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 20 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.
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.

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 capture antibody against Collagen Type VI Alpha 1 (COL6a1). After incubation and washing, the assay 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 Collagen Type VI Alpha 1 (COL6a1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration.
Source Mouse
Synonym Mouse COL6a1 ELISA Kitt
Detection Type Double antibody sandwich method
Composition
Name 9 6 T  match   set remark
Pre-coating 96 Well plate 8 Hole ×12 Strip without
Standard 2 branch
Dilute as per instructions
Universal diluent
2×20mL
without
Concentrated biotinylated antibody ( 100× )  
120uL
Dilute as per instructions
Concentrated enzyme conjugate ( 100× )
120uL
Dilute as per instructions
20× Washing liquid
2×10mL
Dilute as per instructions
Bottom thing ( TMB )
10mL
without
Stop liquid
6mL
without
Sealing film
4 Zhang
without
Instructions
1 Share
without
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 0.62-40 ng/mL
Applications Tissue homogenates, cell lysates, and other biological fluids
Shipping Notes
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Exchange/Return Notes
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SKU: 64466293908

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Color: 01 - Light Grey, Size: Queen
These sheets! I love them. They are so soft and comfortable to sleep on. The quality of my sleep has improved dramatically since I bought these sheets. I tend to sleep hot, yet I stayed at a comfortable temperature all night with these sheets. Our bed takes a 21” fitted sheet pocket, and this is the first fitted sheet that I’ve found that actually covers the corners, with a little extra to spare. I will be ordering these again! The price is great, the fit is great, the colors are great, the comfort is great. What’s left to say, except—they’re great!
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Color: 02 - White, Size: Queen, Color: 02 - White, Size: Queen
I was pleasantly surprised by the quality of these Bedsure sheets. They feel lightweight yet durable, offering a soft and cooling touch that’s perfect for warm nights. The deep pockets fit my thick mattress securely without slipping off, which is a big plus. After several washes, there’s been no pilling, and the color stayed bright white without fading or yellowing. For the price, the value for money is excellent — they feel as good as some luxury brands I’ve tried. Overall, a great combination of comfort, durability, and quality. I’ll definitely be buying another set in a different color soon!
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Color: 01 - Light Grey, Size: Queen
It should be 4.5 stars because these sheets are great but I got the light grey and it was more of a barely not white. I would have liked it to be a bit more true to the color choice. That aside, they have been through 6 washes with minimal to no shrinking (washed on hot and dried as hot as my dryer allows but it’s pretty sensitive so it shuts off as soon as it senses dry. Other machines may vary.) I like how the sheet is labeled with a “top or bottom” tag. I’ve experienced no slipping and so far they show no wear and tear like some similar sheet sets I’ve purchased. I would recommend this at the price. They feel soft and have proven durable. I like to change my bedding style a lot and appreciated the wide selection of colors. Again, this was not true to color (very light grey, almost white, when I’d ordered something visibly much darker in the image but called light grey), but outside of that it was a great set right out of the package.
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Color: 01 - Light Grey, Size: Queen, Color: 01 - Light Grey, Size: Queen
I've been having trouble finding sheets that didn't pill constantly. THESE HAVENT. The texture of these is so comfortable and it keeps me nice and cool at night. The straps underneath actually help them stay on the bed with no falling off!! I have a queen size bed and it's just me and I didn't have any problems getting these on to the bed either. They are extremely good quality for the price. The sheets I had before this were juicy couture and they pilled so bad they are garbage after 90 days. I will continue to purchase bedsure sheets from now on!!
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