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Description
Rat CCR2 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 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 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 Chemokine CC-Motif Receptor 2 (CCR2) 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 Chemokine CC-Motif Receptor 2 (CCR2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Chemokine C-C-Motif Receptor 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | C-C chemokine receptor 2 (CCR2 or CD192 (cluster of differentiation 192)) is a protein encoded by the CCR2 gene. CCR2 is a C-C chemokine receptor. The CCR2 gene is located in the chemokine receptor gene cluster region. It expresses two alternatively spliced transcript variants. This gene encodes two isoforms of the receptor for monocyte-inducing protein-1 (CCL2), a chemokine that specifically mediates monocyte chemotaxis. The receptor encoded by this gene mediates agonist-dependent calcium mobilization and adenylate cyclase inhibition. | |||||||||||||||||||||||||||||||||
| 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 homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.2 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 3
I don't know what it's supposed to be like
I have never had Biltong so I have no idea how this compares to others. I was pretty excited about it because it claims to be a tender beef snack, better than jerky. In my experience though, there were a couple things I should note.
It's not that tender. It's not super chewy like some very dry jerky is but it's sort of like a very thick paper. I think it would be a really good texture if it were more tender but as-is, I don't know if people purely prefer it or if it's sort of a traditional thing that has a texture they try to nail.
It's not big on flavor. Clearly it is spiced for flavor to a degree but it's very muted. Regarding heat, I have a lot of spicy food so maybe people that don't would think otherwise but, to me, calling it spicy is a huge overstatement.
I need to try other biltong because it's entirely possible that this is a prime example and I just don't know it, but if it is I can't say I'm a big fan.
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Reviewed in the United States on March 29, 2019
★★★★★ 5
Good Cut/Texture, Spicy Flavor Isn't the Best - Try Traditional!
Flavor Name: Spicy, Size: 4 Ounce (Pack of 1)
Update: Great customer service! They provided a pack of traditional flavor after seeing my review (originally 3 stars, now 5). The traditional flavor is MUCH better and passed the taste-test from everyone in my house. I imagined traditional flavor would be like spicy - just milder. I was wrong, it's a very different flavor, more familiar to American jerky, but still very unique and delicious. Combined with the unique texture (biltong is new to me), it's hard not to eat this by the handful. Getting the right flavor completely changed my opinion. Try traditional first!
Original Review: I appreciate the nutrition, great for snacking. The cut/texture is good - seems like thicker cuts of dried meat sliced wafer thin. I tried Spicy Biltong, which wasn't great. I normally share a little bit of jerky with my young son and even my wife has some occasionally. We all agreed that the flavor wasn't great. Maybe other flavors are better. Better flavor would have made all the difference, but at this price, I probably won't risk it.
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Reviewed in the United States on February 6, 2019
★★★★★ 1
Buy if you like shredded meat I guess
Flavor Name: Spicy, Size: 2 Ounce (Pack of 1)
I was so excited to try these “slices” of beef because I grew up eating deer biltong made from deers my uncles would go hunt. Some of the best snacks I ever had, absolutely delicious. So when I decided to see if any biltong was actually sold around here I was surprised and pleased this was available. Unlike beef jerky which uses lots of sugar for some reason I was hoping this would have more meaty flavor but it doesn’t really. Although that might be due to the fact that I did get “slices” of meat, no what I got was basically shredded and powdered. It’s salty, but has absolutely no spice whatsoever and is just not good at all.
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Reviewed in the United States on September 28, 2025
★★★★★ 2
Very greasy biltong and not spicy at all
Flavor Name: Spicy, Size: 4 Ounce (Pack of 1), Flavor Name: Spicy, Size: 4 Ounce (Pack of 1)
The flavor is ok but the spice is almost nonexistent. It’s super greasy/oily, with big pieces of fat, so if you’re expecting a nice lean, dry, and spicy biltong, this is not the right one to try. I took the photo to try and capture the shiny, wet, translucent appearance of this biltong but it’s hard to see. Maybe it’s just personal preference but I’m used to biltong being dry and opaque with nearly zero fat chunks and not leaving a film of oil on my hands after eating it.
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Reviewed in the United States on July 6, 2025
★★★★★ 5
Most like South African Biltong
Flavor Name: Spicy, Size: 4 Ounce (Pack of 1)
Firstly, this is NOT JERKY!! So please stop comparing it to that disgusting, over-processed, artificially flavored stuff. Nothing can compare to good biltong. It is seasoned and air dried meat that is the most amazing high protein snack available. Jerky doesn't deserve to be in the same room as biltong, let alone compared to it.
I am a South African recently relocated to the USA and have been trying various Biltong brands available. This is by far the best. I prefer the spicy to the original, but that it a personal preference, the original is also very good, but could do with a bit more flavor. The dry wors is also good, but I am a big biltong eater and always have been. For those of you who don't know, dry wors is a thin sausage (sausage = wors), that has been air dried. It is not a brat type sausage that most Americans are familiar with, it is more a kielbasa or Argentinian style sausage (but not quite).
Thank you, being able to have this, has prevented me from going crazy some days. Now I just need to try your boerewors!
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Reviewed in the United States on September 16, 2021