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Description
Human RAB5C ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. 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 the Ras-related protein Rab-5C (RAB5C). 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-5C (RAB5C) 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-5C ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | RAB5C (RAB5C), a member of the RAS oncogene family, also known as Ras-associated protein Rab-5C, is a protein encoded by the RAB5C gene. It belongs to the Ras-associated protein family and is a small GTPase in the Ras superfamily. It is believed to ensure the fidelity of vesicle docking and/or fusion with its correct receptor compartment. It regulates Rac-mediated cell motility and is a key regulator of the trafficking of the CD93/Multimerin-2/β1 integrin complex in endothelial cell adhesion and migration. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenates and other biological fluids |
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4.8 ★★★★★
Based on 18 reviews
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Product Reviews
★★★★★ 3
They are using different/cheaper shredded foam then they were 1 year ago
Color: Memory Pillow, Size: Standard, Color: Memory Pillow, Size: Standard
The shredded foam used last year was really nice and soft and even firm to a point where your head is supported perfectly while you sleep. now the 2 extra pillows I received this weekend have completely different shredded foam which seems to be cheaper. Also when you unzip the pillow cover and get to the part where you can unzip the other cover where the foam is no longer is there. so they are using cheaper shredded foam and a cheaper cover to where the foam is and you can no longer unzip the second cover to see the foam. but I can easily tell that they are using different and cheaper foam which isn't true memory foam. their new blend is too fluffy and the pillows are floppy and too soft. the old ones i bought 1 year ago are heavy pillows that are true memory foam which your head molds into perfectly and are amazing. these new ones they are making aren't worth it. In one picture you can see the cover with different colored cheap foam and the other picture with the green and white thick premium quality shredded memory foam. The other pictures show how the old ones are thick and firm and moldable for you head and the other 2 are floppy because they use cheaper shredded foam. They need to use the shredded foam they used last year! it was really good! Now they are not true memory foam pillows anymore due to changing from premium shredded foam to cheap shredded foam.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 18, 2024
★★★★★ 5
Great comfortable, pillows.
Color: Black(4pack), Size: Standard
These are some great, comfortable pillows. They are soft and feel great. They are good for sleeping on or just using them to be comfortable in bed.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 23, 2026
★★★★★ 5
Enjoyable
Color: Grey, Size: Queen
Holds their shape and stay cool.
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Reviewed in the United States on May 24, 2026
★★★★★ 5
So comfy
Size: King (Pack of 1), Style: Standard Loft (4" - 5")
I spent time looking up/watching videos on pillows for neck pain and this one comes up a lot. I’m someone that used to wake up every few weeks with a stiff neck so bad it could make me cry at work. I haven’t had that happen once after months of using this pillow. I look forward to laying my head down on it every night. It’s a luxury experience every time. I think what it does so well is maintain its shape. You can really squish it up how you want and it stays there even after hours. Moldable but somehow still firm. It’s expensive but to me the value is there because I no longer get crazy neck pain from sleep. Worth trying it out for sure. Btw I was stuck between this is and Purple brands couple top end options. I never got around to trying them because I tried this one first and was very satisfied!
Feb 2026. Still use it every night and it's still protecting me from neck pain I used to get from sleeping. It's held up amazing. Hope to use it for many more years.
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Reviewed in the United States on January 18, 2025
★★★★★ 5
Saatva Pillow is great
Size: Standard/Queen (Pack of 1), Style: Standard Loft (4" - 5")
So far this is the best pillow I have had in the last 10 years and I have tried two Purple pillows a couple different memory foam and Gel pillows at least 4 more pillows that I can't even remember the names of. My neck and shoulders are almost pain free. I have also been doing more stretching of my arms which is also helping, but this pillow has really made the most difference in the pain I have had, I felt difference from the first time I put my head on it. It's comfortable, the perfect height I got the 4"-5", I also got the queen size and it's great. It supports my head just right. I also ordered the Saatva Classic 14" mattress from the company and I sure hope it's as good or better would be great because I have back issues for the last 50 years.
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Reviewed in the United States on September 16, 2025
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