SKU: 76316404615

Rat IL12B ELISA Kit

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Description

Rat IL12B 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. 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.

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.

Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis.

Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles.

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 200 pg/mL). Then dilute to the following concentrations: 200 pg/mL, 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 200pg/mL standard working solution into the first EP tube and mix thoroughly to make a 100pg/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 utilizes 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 Interleukin 12B (IL12B) capture antibody. 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 Interleukin 12B (IL12B) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration.
Source Rat
Synonym Rat Interleukin 12B ELISA Kit
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
Background The beta subunit of interleukin-12 (also known as IL-12B, natural killer cell-stimulating factor, cytotoxic lymphocyte maturation factor p40, or interleukin-12 subunit p40) is a protein encoded by the IL12B gene. IL-12B is a common subunit of interleukin-12 and interleukin-23. This gene encodes a subunit of interleukin-12, a cytokine that acts on T cells and natural killer cells and exhibits a wide range of biological activities. Interleukin-12 is a disulfide-linked isoform composed of a 40 kD cytokine receptor-like subunit encoded by this gene and a 35 kD subunit encoded by IL12A. This cytokine is expressed by activated macrophages and is an important inducer of Th1 cell development.
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 3.12-200 pg/mL
Applications Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids
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SKU: 76316404615

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4.8 ★★★★★
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Amazon Customer
San Leandro, US
★★★★★ 1
They are hard to chew
Gummies are old and hard, still tried them cause I want a fantastic kitty and it actually did the opposite. It’s been itchy and irritated with every that comes in contact with it. I never had a problem down there but now I do. It’s a no go for me.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 3, 2026
J
jwilt8184
Pawtucket, US
★★★★★ 5
Tasty, Easy‑to‑Take Gummies for Daily Feminine Balance Support
I picked these up because I wanted a simple, vegan option to support pH balance and overall feminine wellness, and these gummies have been really easy to add into my routine. The Hawaiian pineapple flavor is surprisingly good—sweet without being artificial—and the texture is soft and easy to chew. I like that they combine multiple areas of support in one: healthy odor, flora balance, and immune support. It makes them feel like a well‑rounded daily supplement instead of juggling multiple bottles. They’re also non‑GMO and vegan, which is a big plus if you’re trying to keep your routine clean.
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Reviewed in the United States on April 16, 2026
C
CEAM
Whiting, US
★★★★★ 4
Tastes Great, But More General Wellness Than Targeted Feminine Support
I tend to prefer gummies over capsules whenever possible, so these Yalei 4-in-1 Nuora Feminine Balance Gummies caught my attention as a potential replacement for my usual women’s probiotic. The bottle contains 60 gummies (30 servings), with a formula centered around vitamin C, pineapple fruit powder, and Bacillus coagulans (1 billion CFU). The ingredient list is clean and clearly labeled, and I appreciated that it’s free from common allergens like soy, dairy, gluten, and tree nuts. The outer and inner tamper seals were intact. The manufacturing and expiration dates were also easy to read, which is something I always look for with supplements. Taste-wise, these are genuinely enjoyable. The pineapple flavor is pleasant, not overly artificial, and there’s no lingering aftertaste. The texture is firm without being sticky, which makes them easy to take daily. Honestly, these fall into that category where it’s tempting to eat more than the serving size. Where things became more nuanced for me was performance. I switched from a more targeted women’s probiotic that included specific Lactobacillus strains and additional D-mannose and cranberry support ingredients. Within about two weeks of making the swap, I experienced a UTI flare-up, which led me to go back to my previous formula. That experience made the difference in formulation stand out. My prior supplement included strains commonly associated with vaginal microbiome support along with additional complementary ingredients, whereas this product uses a single Bacillus-based probiotic strain. From my experience, this felt more like a general wellness probiotic gummy rather than something formulated for more targeted feminine health concerns. That doesn’t make it a bad product–it just depends on what you’re looking for. If your goal is a simple, good-tasting daily probiotic with light supportive ingredients, this fits that role well. If you’re specifically trying to maintain balance in more sensitive or recurring situations, you may want something more specialized. The brand also includes details like GMP manufacturing claims and batch testing which adds a layer of reassurance, though it does appear to be a newer brand. At its current lower price point, I do think it offers decent value for a general-use gummy. For me personally, though, I’ll be sticking with a more targeted formula moving forward.
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Reviewed in the United States on March 17, 2026
B
Brian J.
Alexandria, US
★★★★★ 5
Probiotic fruit snacks
Disclaimer: vine review of an item I requested free* These taste great, just like fruit snacks. I've gotten a lot of gummies from this brand and haven't found a bad one yet. Pineapple isn't even a flavor I usually like, but these taste good. Somehow they're always sweet with no weird aftertaste without having added sugar. Not too sticky, it doesn't leave residue on your fingers, but they do sometimes stick together. Mainly I got these to have some fruit snacks to eat, but the probiotics are a nice bonus and now I can pretend I'm being healthy when I eat my candy.
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Reviewed in the United States on April 4, 2026
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Verified Purchase
Daissy strizich
Grantham, US
★★★★★ 5
Actually Helped My Metabolism and Gut Health!
I’ve been taking these capsules for about two weeks now and honestly I can feel a noticeable difference. My metabolism used to feel very slow, and I often felt bloated and heavy after eating, but since starting this supplement my digestion feels much better and my stomach feels lighter. I also noticed I have more energy throughout the day and less swelling. The capsules are easy to take and gentle on my stomach, which is important because some gut health products can feel too strong. I like that this formula includes probiotics and berberine together because it feels like it supports overall wellness, not just digestion. Of course everyone’s body is different, but for me this product has been working very well so far. I’m happy with the results and will continue taking it.
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Reviewed in the United States on May 20, 2026

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