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Description
Rat RF 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: 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: 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. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant and centrifuge 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. Cell Supernatant: 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., 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 the 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. Preparation before testing: 1. Please remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 1000 mIU/mL). Then dilute according to the following concentrations: 1000 mIU/mL, 500 mIU/mL, 250 mIU/mL, 125 mIU/mL, 62.5 mIU/mL, 31.25 mIU/mL, 15.625 mIU/mL, and 0 mIU/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 1000 mIU/mL standard working solution into the first EP tube and mix thoroughly to make a 500 mIU/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 enzyme conjugate are sequentially added to microwells pre-coated with rheumatoid factor (RF) capture antibodies. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of rheumatoid factor (RF) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat rheumatoid factor ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Rheumatoid factor (RF) was the first autoantibody discovered in rheumatoid arthritis. It is defined as an antibody targeting the Fc portion of IgG, with different RFs recognizing different parts of the IgG-Fc. RF and IgG bind to form immune complexes, contributing to the disease process. Rheumatoid factor can also be a cryoglobulin (an antibody that precipitates after cooling a blood sample) and can be either type 2 (monoclonal IgM to polyclonal IgG) or type 3 (polyclonal IgM to polyclonal IgG). Although IgM is predominant, rheumatoid factor can be any immunoglobulin isotype, namely IgA, IgG, IgM, IgE, and IgD. | |||||||||||||||||||||||||||||||||
| 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 | 15.6-1000 mIU/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, cell supernatant, tissue homogenate, etc. |
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4.5 ★★★★★
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Product Reviews
★★★★★ 5
WorthIt! Nighttime ritual , Creates functional system. We drink on evenings of heavy dinners
Flavor Name: Unflavored, Size: 1.1 Pound (Pack of 1)
WorthIt! Every other night before bed routine. Creates a happy and well functioning system. We add a splash of flavored electrolytes . Nice treat as well.
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Reviewed in the United States on May 18, 2026
★★★★★ 4
Good quality
Flavor Name: Unflavored, Size: 1.1 Pound (Pack of 1)
Seems to work as expected and looks consistent in quality. The only problem I have with it, is the opening being too small for my hand when it is running low. I bought a another brand last time with the same quality and I could reach inside. I comfortably fit men's large size gloves, so my hands aren't huge. Maybe a long handle scoop would help...
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Reviewed in the United States on April 27, 2026
★★★★★ 5
No taste or texture - it completely dissolves!
Flavor Name: Unflavored, Size: 1.1 Pound (Pack of 1)
Easy way to add fiber to diet. Stir it in coffee or a non-carbonated cold drink. You can't tell it as even added because it completely dissolves.
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Keeps me feeling full. Perfect addition to protein shake
Flavor Name: Unflavored, Size: 1.1 Pound (Pack of 1)
Tasteless pretty much. I’ve added to coffee, lemonade, cereal with milk, sauces, stir fry, pasta sauce, yogurt. It’s great addition to my protein drink. It helps me both keeping good on the bathroom but it helps me feel more full and snack less. It has been a secret in helping me lose weight.
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Reviewed in the United States on April 3, 2026
★★★★★ 5
A Convenient Fiber Supplement- Digestion, Regularity, Microbiome, Satiety, Hair, Skin, Nails
I’ve tried quite a few fiber supplements over the years, and this one stands out mostly because it combines digestive support with beauty-focused ingredients instead of being just another plain fiber gummy. The formula includes inulin and chicory root fiber, along with a fairly large probiotic blend containing multiple strains like Lactobacillus acidophilus, Lactobacillus plantarum, and Bacillus coagulans. That combination makes sense since prebiotic fiber and probiotics are often paired together to help support digestive balance and regularity.
The added hair, skin, and nail ingredients are also interesting here. Biotin is commonly used for supporting stronger nails and healthier-looking hair, while collagen peptides and keratin are popular ingredients for skin elasticity and hair structure. Vitamin C helps support collagen production naturally, and Vitamin E adds antioxidant support. The apple cider vinegar and saw palmetto are smaller additions, but they round out the formula nicely for overall wellness support.
One thing I appreciated is that these are gummies instead of powders or large capsules. Fiber supplements can sometimes feel like a chore, especially the gritty drink mixes, so having a chewable option made it easier to stay consistent. The blueberry flavor was actually pleasant and not overly artificial. Texture-wise, they were softer than expected and easy to chew.
As far as results, I mainly noticed gradual digestive benefits after about a week or so of consistent use. Things felt more regular and less sluggish overall, especially on days where my diet wasn’t as balanced as it should have been. The fiber content is fairly substantial for gummies, so I’d definitely recommend drinking enough water with them because increasing fiber too quickly can sometimes cause bloating at first. Starting with a smaller serving may help if you’re not used to fiber supplements.
I can’t say I noticed dramatic changes with hair, skin, or nails during the time I used them, but those ingredients usually take longer to show visible effects anyway. Even without obvious cosmetic changes, the combination of biotin, collagen, keratin, and antioxidants gives the formula a more well-rounded feel compared to standard digestive gummies.
Another positive is the allergen-friendly formula. Being gluten-free and free from dairy, wheat, peanuts, and eggs makes it accessible for many people with dietary restrictions. The overall ingredient profile feels thoughtfully put together instead of overloaded with unnecessary fillers.
Overall, I think this works best for someone looking for an easy daily fiber supplement that also offers some additional wellness support beyond digestion alone. The biggest benefit for me was the digestive regularity and convenience factor. The beauty-support ingredients are a nice bonus, even if they may require long-term consistent use before noticeable results appear.
**As always, results can vary from person to person. Consulting a healthcare provider before starting any new supplement is advisable. Keep well (:
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Reviewed in the United States on May 11, 2026