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Description
Rat LC3 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 frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the 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 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 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 Microtubule Associated Protein 1 Light Chain 3 (LC3). 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 Microtubule Associated Protein 1 Light Chain 3 (LC3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Microtubule Associated Protein 1 Light Chain 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Microtubule-associated protein light chain 3 (MAP1LC3), often abbreviated as LC3, is a soluble protein with a molecular weight of approximately 17 kDa. LC3 is ubiquitous in mammalian tissues and cultured cells and is a key component of autophagy, the recycling system of eukaryotic cells. It is incorporated into the inner and outer membranes of autophagosomes during autophagosome biogenesis. Therefore, LC3 is a specific marker of autophagy, particularly autophagosome formation. | |||||||||||||||||||||||||||||||||
| 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, cell lysates, cell culture supernatants, and other biological fluids |
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4.6 ★★★★★
Based on 22 reviews
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Product Reviews
★★★★★ 4
good spray, but strong alcohol smell
Style: SPF 50, Size: 5 Fl Oz (Pack of 1)
I like Alba Botanica, but 3.5 starts since:
- the spray goes way too fast (that can be true for all continuous sprays). I used it 4 - 5 times and it seems I have very little left.
- lots of waste when spraying... probably is best to spray your palm and then massage it on your body
- once it's sprayed it smells like alcohol, alot!!! Other brands I tried don't seem to smell so much of alcohol.
- My kid developed strange rash once it was applied, not red dots, but white bumps all over the arms and legs, cannot say its from this spray, but I'm a bit suspicious.
The good party it's it is not sticky or greasy.
Smells good (after alcohol evaporates).
I used it a couple of times, applied once only and stay several hours under the sun and I was fine, no sunburn.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 20, 2016
★★★★★ 5
Love it!
Style: SPF 50, Size: 5 Fl Oz (Pack of 1)
Love this stuff and it's cruelty free!!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 4, 2026
★★★★★ 1
Alba packaging stinks
Style: SPF 50, Size: 5 Fl Oz (Pack of 1)
Complete waste of money! I ended up tossing it out because it’s IMPOSSIBLE to use. You should be able to twist the top to lock/unlock and spray it. NOPE. The top just spins and doesn’t catch or you cannot twist it at all. It’s also virtually impossible to spray. I had to hold it against my torso and press with both thumbs to get it to spray. Meanwhile, it’s leaking, the bottle is getting slippery. And then the spray goes EVERYWHERE, it’s not a narrow spray but it’s as if you’re dusting crops. I tried to see if it needed a certain pressure point, two fingers, bloody hell it’s not a Magic Mouse you should be able to unlock and spray it with one hand.
Anyway, Alba needs to do some QC on their packaging. The last pump lotion I got, the pump also did not work. It just spun and spun and could not be unlocked. I had to wash and reuse the pump from an old bottle.
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Reviewed in the United States on June 21, 2024
★★★★★ 5
A+
Style: SPF 50, Size: 5 Fl Oz (Pack of 1)
Best, safest sunscreen!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 4, 2026
★★★★★ 5
Best sunscreen brand! Fair, sensitive-skinned and rely on this
Style: SPF 50, Size: 5 Fl Oz (Pack of 1)
Alba Botanica is my favorite brand for sunscreen products and it checks all the boxes on my list: Excellent long lasting coverage, fine mist spray and easy coverage cream versions, great for my sensitive skin (no breakouts- I do NOT use it on my face though), the Hawaiian fragrance is pleasant, it’s reef safe (a big one for me as a snorkel/dive aficionado), and organic ingredients. The large size lasts 5 full days when on a boat all day in the Carribbean, and I only apply it twice (once in morning, and again midday.) I have fair skin, and I never burn. The spray version is, by nature of its aerosol capacity, a bit drying on the skin, but this is to be expected, and it creates a long lasting barrier. The cream version is a wonderful emollient, although not quite as long lasting. Best deal is only in the summer season at CostCo for a 2 pack... in my searches, Amazon has the best deal otherwise.
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Reviewed in the United States on January 28, 2021
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