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Description
Human RAB5A 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: 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 freezing and thawing 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 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 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. 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 directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 the Ras-related protein Rab-5A (RAB5A). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Ras-related protein Rab-5A (RAB5A) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Ras-related protein Rab-5A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | RAB5a, also known as RAB5a, a member of the RAS oncogene family, is a protein encoded by the RAB5a gene. It is localized to early endosomes, where it participates in the recruitment of RAB7A and the maturation of these compartments into late endosomes. It drives endosomal maturation by transporting vacuolar (H+)-ATPase (V-ATPase) from the trans-Golgi network to endocytic vesicles. It is required for fusion of the plasma membrane and early endosomes and helps regulate the extension of filaments. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.7 ★★★★★
Based on 176 reviews
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Product Reviews
★★★★★ 5
Pleasant Scent, Effective Cleaner
Scent: Lemon Verbena Concentrate, Size: 32 Fl Oz (Pack of 2)
I love the delicate verbena scent in Mrs. Meyers Clean Day Multi-Surface Cleaner. It’s very mild compared to other common household cleaner’s like Mr. Clean (Mr. Clean is one that knocks me over and I can’t stand the smell). Many years ago it was fine, but the scent has changed and become stronger, and NOT for the better. Manufacturers on the whole, use way too much scent in their products, whether it’s laundry detergent, fabric softener, hygiene products and anything else that scent is added to. Just take a walk down the detergent isle of your grocery store and if you aren’t immune to scent, you will find it overwhelming. This cleaner is not overpowering and smells just like verbena, a fresh, clean, green smell. I also love the way it cleans my wood floors, sinks, countertops, stainless steel fridge, stove & dishwasher, & everything else I clean with it. It leaves the surfaces “squeaky clean” without any streaks. Is it a good deep cleaning cleaner? I don’t know for sure since I’ve never used it to clean up muddy, greasy, grimy, dirty areas in my home. It does remove dark marks on painted walls but I haven’t tried it on heavy dirt and grime. I imagine it’s comparable to Murphy's Oil Soap, which is another brand I love and have used for years. If the scent of cleaners is an issue for you, and you like verbena, you should love this cleaner. A little bit goes a long way in a bucket of water (for washing floors, etc.). I usually use allot less than is recommended (in everything from laundry detergent to household cleaners). Especially my laundry detergent. I used to use the recommended amount and found with concentrated products, the amount recommended is way too much. I could feel the soap in my clothing after the wash cycle finished. I had to re-rinse all my loads. Then I got smart and cut way down on detergent and included vinegar in the final rinse cycle. Manufacturers love to sell more product, so don’t go by what they state on the container. Use your own wise judgement. On a side note, I love Costco’s Industrial Strength powdered laundry detergent but I cut way back to 1/4 cup per med to large loads. The white dress shirts and tee shirts are much brighter & whiter compared with many other detergents I’ve used in the past. I highly recommend this product for all of the above reasons. Give it a try at least once. You may love it too!
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Reviewed in the United States on June 17, 2024
★★★★★ 5
Excellent cleaning product, but lavender is a bit too strong for me.
Scent: Lavender Spray, Size: 16 Fl Oz (Pack of 3)
I bought 3 bottles of the lavender cleaner, and have used it to clean a number of different surfaces. In one of the bathrooms, I used it to clean dust from 6-panel doors, the cabinets, the counter and sink, the inside and outside of the tub area, and the outside of the toilet. When I was done, everything in the bathroom looked so clean. Even the dust that collects in the corners of the 6-panel doors was gone.
Next, I tried to clean stainless steel refrigerator in the kitchen. The refrigerator wasn't dirty, but it had streaks from water dripped from the ice maker and fingerprints. This cleaner didn't work as well on the refrigerator. Although it did remove the fingerprints and water streaks, it left streaks that I had to clean using another method. Actually, a clean washcloth and warm water worked very well to remove the streaks. This cleaner also left streaks on my built in stove that has a glass front. So, I don't believe this cleaner works well on reflective surfaces such as mirrors, glass, or stainless steel. Although it does clean these surfaces well, it leaves a residue that causes streaks.
The last surface I tried cleaning today was some of my kitchen cabinets. These cabinets collect not only anything in the air from cooking, but, since I am a smoker, they also collect some nicotine. On the cabinets that I cleaned, this cleaner did remove stains from both as was evidenced by the washrags I was using. To-date, the only cleaner I have found that would effectively clean these stains was Windex. So, I was impressed at how well this cleaner worked. It was working so well, I wanted to keep going, but, unfortunately, ran out of time.
Overall, I would recommend this cleaner for anything but mirrors, glass, and stainless steel. The lavender scent of the bottles I got was a nice scent, but a bit overwhelming in my opinion. Once I use up the bottles that I have, I will be trying one of the other scents, possibly the honeysuckle or lemon scents.
UPDATE 05/21/25:
We don't use our dining room which is furnished with a 40 year old solid wood dining room set. All too often the table is used for keeping appliances or other items that I plant to get rid of or need to find a permanent storage place. I finally got the table clear off and saw a decent layer of dust on it. So, I decided to try this cleaner on it. This cleaner not only removed all dust and spots on the table top but also the layers of old furniture polish. I was so pleased that the table was clean with a natural shine that did not feel oily. Now all I have to do is clean the rest of dining room furniture. I will definitely be using this cleaner.
I hope this review was helpful for you.
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Reviewed in the United States on April 23, 2025
★★★★★ 4
Smells amazing while cleaning, light lasting fragrance
Scent: Peppermint, Size: 16 Fl Oz (Pack of 1), Scent: Peppermint, Size: 16 Fl Oz (Pack of 1)
I really enjoyed using this Mrs. Meyer’s Clean Day Multi-Surface Cleaner, especially because of the peppermint scent. It smells incredibly fresh and pleasant while you’re cleaning, not overpowering, just very clean and uplifting.
I tested it on multiple surfaces including countertops, tables, and other everyday areas, and it performed well across the board. It cleans effectively without leaving residue or streaks, and it feels gentle enough for regular use.
The only reason I’m giving it four stars instead of five is because the scent fades fairly quickly after cleaning. While it smells amazing at first, the fragrance doesn’t linger for very long. That may be a plus for people sensitive to strong smells, but if you’re looking for a long-lasting scent, this might fall a bit short.
Overall, it’s a solid multi-surface cleaner with a beautiful scent and reliable cleaning performance. I’d definitely repurchase, especially during seasonal releases.
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Reviewed in the United States on February 7, 2026
★★★★★ 5
Bang for your buck
Scent: Lemon Verbena Concentrate, Size: 32 Fl Oz (Pack of 2), Scent: Lemon Verbena Concentrate, Size: 32 Fl Oz (Pack of 2)
The amount of spray bottles you get from this concentrate is actually insane. I won’t have to buy surface cleaner until next year. No but seriously, this is a great price point and you get so much from it. All of the Mrs Meyers products have awesome conscious ingredients that I feel comfortable having around my kids and pets (and they’re insanely gross creatures too) The scent is just a cherry on top!!
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Reviewed in the United States on May 26, 2026
★★★★★ 5
a miracle in a jar
Scent: Earthy, Nutty, Refreshing, Size: 4.06 Fl Oz (Pack of 1)
This item did exactly what it was expected to do, and I couldn’t be happier with my purchase. It works as described, is easy to use, and the overall quality exceeded my expectations.
The price was great for what you get, making it an excellent value. Everything about it—from performance to convenience—has been spot on. My sister has noticed an extreme growth in her hair since using this product.
I’m very satisfied and would definitely recommend this to anyone looking for something reliable and well worth the money!
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Reviewed in the United States on May 5, 2026