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Description
Mouse PKCη 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 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 Protein Kinase C Eta (PKCη). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Protein Kinase C Eta (PKCη) Eta in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Protein Kinase C Eta ELISA Kit Eta | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Protein kinase Cη (PKCη) is an enzyme encoded by the PRKCH gene. Protein kinase C (PKC) is a family of serine- and threonine-specific protein kinases that are activated by calcium and the second messenger diacylglycerol. PKC family members phosphorylate a variety of protein targets and are known to participate in numerous cellular signaling pathways. PKC family members are also the primary receptors for sorbitol esters, a class of tumor promoters. Each member of the PKC family has a specific expression pattern and is believed to play a distinct role in the cell. The protein encoded by this gene is a member of the PKC family. It is a calcium-independent and phospholipid-dependent protein kinase. It is primarily expressed in epithelial tissues and has been shown to reside exclusively in the nucleus. This protein kinase regulates keratinocyte differentiation by activating a protein kinase cascade that activates the MAP kinase MAPK13 (p38delta) that targets CCAAT/enhancer binding protein α (CEBPA). It has also been shown to mediate transcriptional activation of the transglutaminase 1 (TGM1) gene. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.6 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 5
Flavorful
Flavor Name: Traditional, Size: 2 Ounce (Pack of 3)
Best jerky ever I love everything about it!
Jeff Johnson
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 3, 2026
★★★★★ 5
Excellent flavor and texture to this jerky with no sugar makes it my new favorite
Flavor Name: Traditional, Size: 4 Ounce (Pack of 1)
I have recently adopted a low carb, high protein diet combined with weight training, so I'm always looking for new sources of protein. I use protein powder as little as possible, and eat more from natural sources.
I have a brand of beef jerky I really like, but traditional beef jerkies have a lot of sugar (10g protein to 6g of sugar). Biltong, however, has more protein per oz (16g) and 0g of carbs or sugar. Simple ingredients, and nothing funky. So a big win on the nutrition front.
That is great, but what won me over was the flavor. It has a nice hearty flavor, and has a little wetness to it that makes the texture nice to chew. I have tried another brand of biltong, but I found it too dry for my taste (kinda of like chewing on plastic). So another win on flavor and texture.
Pricing is okay. Good beef jerky is not cheap, so you get what you pay for. I would like to buy in bulk, so bigger packages with better discounts would be a good offer I would jump on immediately.
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Reviewed in the United States on May 22, 2019
★★★★★ 5
Good Product
Size: 3 Panel-102'', Color: Beige
I got these dividers for outdoor use for some privacy from neighbors. They arrived quickly and were easy to set up. They look good, and though the legs don’t offer stability against any amount of wind—which I expected as it wasn’t advertised for outdoor use—all it took was placing a cinder block on the feet to hold it up right. Coverage is good, it’s easy to fold and move as needed, and it’s light weight. All in all a decent product. I will caution the purchase of “used like-new,” as both of the dividers I ordered were previously returned items, and one came missing a cap, instructions were not included, and the poles were all mixed up and not properly labeled. If I had not ordered a second divider (which did include the instructions) I would have had a much harder time figuring out which pieces were which and how to put them together.
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Reviewed in the United States on May 30, 2025
★★★★★ 5
So versatile! Perfect solution for dividing a room.
Size: 4 Panel-88'', Color: Grey, Size: 4 Panel-88'', Color: Grey
Perfect solution for my space. My office has to double as the hangout room for the kids and I wanted something that would block off my computer so it is left alone. This space allows me to close off my computer as much or as little as I want. It is lightweight yet has sturdy feet. This is important so that if it gets bumped it won’t topple over. The price was great and the build time wasn’t too bad! I would recommend two people for set up. I went with the grey panels and I think I’ll end up hanging some twinkle lights at some point. Great purchase!
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Reviewed in the United States on October 9, 2025
★★★★★ 4
4/5 Stars
Size: 4 Panel-88'', Color: Black
Very tall and easy to set up, the only thing I will say is that they do have large gaps in between, so you won't have 100% privacy unless you add a curtain in the back. Other than that, they are easy to fold, sturdy, and esy to assemble. The gaps are an easy fix so I would say they are worth it.
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Reviewed in the United States on April 16, 2026
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