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Description
Human BLK 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 tyrosine-protein kinase BLK (BLK) capture antibody. 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 tyrosine-protein kinase BLK (BLK) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Tyrosine-protein kinase BLK ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Blk, also known as the tyrosine protein kinase BLK (B lymphocyte kinase), is encoded by the Blk gene. It is a non-receptor tyrosine kinase belonging to the Src family of tyrosine kinases and is involved in B lymphocyte development, differentiation, and signaling. This protein plays a role in B cell receptor signaling and B cell development. It also stimulates insulin synthesis and secretion in response to glucose and enhances the expression of several pancreatic beta cell transcription factors. It utilizes FYN and LYN and plays a crucial role in pre-B cell receptor (pre-BCR)-mediated NF-kappa B activation. It also contributes to BTK activation by indirectly stimulating its intramolecular autophosphorylation. In pancreatic islets, it acts as a regulator of beta cell function by upregulating PDX1 and NKX6-1, subsequently stimulating insulin secretion in response to glucose. It has been shown to interact with UBE3A. | |||||||||||||||||||||||||||||||||
| 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 5 reviews
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Product Reviews
★★★★★ 5
Minty & Whitening Approved!
Size: 4 Ounce (Pack of 4), Set name: Adult Mint
MY FAVORITE fluoride free toothpaste. I still get that slight minty fresh taste and has effectively whitened my teeth as people have noticed and complimented me on.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 1, 2026
★★★★★ 5
Phenomenal NHa toothpaste
Size: 4 Ounce (Pack of 1), Set name: Adult Mint
I am fluoride free. It’s toxic AND did you know it oxidizes the tooth from the inside out and actually MAKES your TEETH YELLOW!? Crest even admitted it. Fluorides risks outweigh the teeth benefits but LUCKILY there’s a more superior solution: nano hydroxyapatite for our teeth remineralization needs. It’s safe and proven to be more effective as a remineralizing agent, unlike fluoride which cannot remineralize on its own - it requires the minerals in your saliva to work, so if your saliva isn’t at the optimal levels, it’s useless to teeth. But I have been on a teeth whitening journey - got my teeth whitened at my dentist but I ate a tangerine and they got stained again. (Stupid me. Bye $325!) While I’ve restored the whiteness back with various at home products, it’s been at the expense of my enamel and has caused a lot of sensitivity. Sensodyne was given to me by my dentist but when I saw it visibly yellow my teeth after the dentist’s first bleaching, I went down a rabbit hole to get to the bottom of it as to why. Additionally, my enamel has been getting more and and more clear, and I needed this to be addressed. NHa has been proven to restore enamel which treats tooth sensitivity instead of just numbing the nerves with potassium nitrate like Sensodyne does which treats the symptom and not the cause (tooth demineralization, sigh… such a western medicine move). When I bought this for the first time I was immediately pleased - there’s NO scary flavor (if you’ve ever tried *natural* toothpastes, especially ones of India, You KNOW!). There’s no cardamom or other cinnamon or out of the ordinary flavors here. The consistency is that of normal toothpaste. Great foaming action. Has helped my sensitivity in 2 applications, I’m 100% without sensitivity but even better, my teeth my are less clear. This truly restores enamel.
But, here’s my gripes. #1 - I have no idea how much hydroxyapatite is in this. I know it works, but as a consumer, I deserve to know what % of the star ingredient is used in a product that I’m specifically buying it for.
#2 - the toothpaste color is… lightish yellow. Think a healthy “mucus yellow” - I’m so sorry for the that description, but when I spit up bits of toothpaste in my sink, it looks like boogers. That’s an issue when I’m busy whitening my teeth at home & I’m avoiding eating foods that stain teeth for at least 2 hours after so that my saliva can naturally seal my dental pores that are being blasted open with peroxide. If I brush my teeth after whitening I’m extremely paranoid that I’m pushing YELLOW toothpaste into my porous teeth, and I don’t need to keep driving myself crazy thinking that the NHa is potentially sealing that color in. I also want to be able to put my NHa toothpaste in clean whitening trays to give myself a mineralizing treatment without worrying about it staining my teeth from prolonged/concentrated contact. Mind you - this is very paranoid of me to think, but I’ve spent a lot fixing my teeth bleaching then restaining fiasco, so I’m not taking any chances. While I don’t think it’s yellowing my teeth at all, because of the color, I’ll never think it’s whitening them.
But don’t let this deter you from purchasing. It helps sensitivity after one use, has shown to make my enamel less clear, and has a normal toothpaste consistency and foams up great. I recommend it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 20, 2024
★★★★★ 5
Great alternative to fluoride toothpaste.
Size: 4 Ounce (Pack of 4), Set name: Adult Mint
Taste great, does the job! Highly recommend!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 29, 2026
★★★★★ 3
No information on active ingredients.
Size: 4 Ounce (Pack of 4), Set name: Adult Mint
Gave 3 starts as an act of goodwill . The toothpaste does not specify the type or percentage of active ingredients it contains. For a beneficial effect, the concentration of nano-hydroxyapatite should be 10% or higher. Based on indirect information, this product appears to contain only about 4.2%.
The manufacturer did not respond to inquiries regarding the active ingredient content, which is concerning.
Additionally, the toothpaste has an unusual taste that is definitely not mint.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 21, 2026
★★★★★ 5
Great for Sensitive Teeth & Leaves My Mouth Feeling Clean
Size: Pack of 1, 4 Ounce, Set name: B. Whitening
Really impressed with the Boka Fluoride Free Toothpaste.
It has a clean, refreshing mint flavor without being overpowering, and it feels much gentler on sensitive teeth than other toothpastes I’ve tried.
My teeth feel smooth and clean after using it, and I like that the ingredients are simple and fluoride free. Definitely something I’ll keep repurchasing.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 25, 2026
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