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Description
Mouse HSPA4 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 heat shock 70 kDa protein 4 (HSPA4). After incubation and washing, the assay 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 heat shock 70 kDa protein 4 (HSPA4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Heat shock 70 kDa protein 4ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | 70kDa heat shock protein 4 (HSPA4) is a protein encoded by the HSPA4 gene. The protein encoded by this gene was originally considered a member of the heat shock protein 70 family. Diseases associated with HSPA4 include Chagas disease and vulvovaginitis. Pathways involved in this gene include the cellular response to heat stress and a potent target for C-MYC transcriptional activation. An important homologue of this gene is HSPH1. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.7 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 3
Style and Length
Size: 38W x 30L, Color: Dark Blue
Pants are not slim fit ,the material is stretchy but if you order them go down to a lower size
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 9, 2026
★★★★★ 5
Sharp and Expensive Quality Tuxedo for special events!!
Size: Small, Color: Black
I researched for several tuxedos for my son's prom, we chose this one, and I'm very glad that we did! The fit is nice, the durability is great. Very classy looking! The quality is really surprising for this price, it looks like a $200 dollar suit. My son is very picky and was very pleased with this tuxedo. The seams are very good and all the buttons are correct. We did receive the tie and everything that was supposed to be with it as I know reviews were saying they didn't get certain things but we did. I really like the adjustable waste because my son is very slim, but tall and has a long torso. This Tuxedo allowed him to be super excited that we have everything we need for his first prom! True to color as we purchased the black and it is sharp!
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Reviewed in the United States on April 17, 2026
★★★★★ 5
Impressive suit for the price
Size: X-Large, Color: Navy Blue
Order suit in my normal size (44R) and it fit perfectly. I’m 6’1” and the pant length set at the correct level. Quality fabric and stitching is well finished. Pants are comfortable at the waist with enough flex to be forgiving.
Very good deal for the price…suit, vest and tie.
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Reviewed in the United States on May 22, 2026
★★★★★ 5
Great for tall, slender teenager!
Size: Small, Color: Light Grey, Size: Small, Color: Light Grey
Finding a suit for a tall, slender teenage boy is about as hard as climbing Mt. Everest. Until I found this one!! I was so pleasantly surprised that this suit fit my 15 year old son like a glove. He is 5'10'' and 140 lbs so he needs pants with the proper length and smaller waist. While the waist was slightly big, he didn't need a belt and the pants fell perfectly with a pair of Nike tennis shoes. I ordered a size small and would say the pants were a 32x32 but waist was probably more like a 31. Jacket and vest were a great fit as well. High quality and easy to steam to get the shipping wrinkles out.
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Reviewed in the United States on May 25, 2026
★★★★★ 4
Fantastic suite for the price!
Size: Large, Color: Khaki
I purchased the gray version to evaluate the suit. I ordered a size up based on other reviews and was pleased to discover everything fit well. The jacket fit well around the arms, chest, shoulders, and even my seemingly ever-growing gut. The vest was the same. The pants also fit well around the waist (thanks to the expanding waistband) butt, and thighs. The only problem was the pants were about 2 sizes too long. Considering the price point, I was felt I was still way ahead after paying to have them altered. After a couple week's worth of wear, I purchased 4 more (gray, black, blue and tan). All fit the same way and were easily altered too. The tailor even had the cashier ask me where I purchased these as he was impressed with the quality.
A couple months in and I'm still pleased, although I've had a few issues. My right pocket on my blue pair has ripped it's seam twice, as has the same pocket on one of my gray pair. Considering it's the same pocket, I would suspect I am just heavy handed but this is not a common problem with any of my other clothes. The pants are a bit on the tighter side when I am sitting down so perhaps the stitching is just not strong enough. I have asked the tailor to give me a super stitch in this area and am just about to pick up the gray pair with this new stitch. I'll try to remember to update later on.
Overall, I feel I have really gotten my money's worth out of these suits, especially considering the prices of others. I have been wearing these daily since June and have gotten many compliments.
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Reviewed in the United States on December 3, 2024
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