Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Aug 27 - Sep 1
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Mouse ATP2B2 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. |
|||||||||||||||||||||||||||||||||
| 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 plasma membrane calcium-transporting ATPase 2 (ATP2B2). 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 plasma membrane calcium-transporting ATPase 2 (ATP2B2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Plasma membrane calcium-transporting ATPase 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Calcium-transporting ATPase B2 (ATP2B2), also known as PMCA2, is an enzyme encoded by the ATP2B2 gene. The protein encoded by this gene belongs to the P-type primary ion-transporting ATPase family, characterized by the formation of an aspartyl phosphate intermediate in the reaction cycle. These enzymes remove divalent calcium ions from eukaryotic cells across very large concentration gradients and play a key role in intracellular calcium homeostasis. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.3 ★★★★★
Based on 19 reviews
Sort
Product Reviews
★★★★★ 4
Beautiful, glorious, and depressing but not without levity
Probably Gilliams most serious film, a story of a dreamer in a dystopian bureaucratic future, plagued by monotony and endless paperwork, our protagonist (played by Jonathan Pryce) isn't sure what he wants in his life, but he knows he wants something different. When the Department of Information Retrieval disappears the wrong person (played by Robert De Niro), our hero goes to reimburse the widow and discovers a woman living in the same building who looks like the woman out of his dreams (played by Kim Greist).
Really stellar performances by Pryce, and a gaggle of supporting actors such as Ian Holm, Michael Palin, Kathrine Helmond, and Bob Hoskins. Greists performance is probably the weakest, but she does enough for the role that the critique of it amounts to a nit pick really.
While this film is considered a cult classic, I would qualify it even less and say its just a classic, full of symbolism with a increasingly repressive tone throughout balanced by Gilliams puncturing, dark humor.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 29, 2024
★★★★★ 5
Classic Terry Gilliam- Watch It!
One of the best Terry Gilliam movies out there. Made in the 1980's, the cast is superb, and it holds up well 40 years on. WAtch it- it's a trip!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 27, 2026
★★★★★ 5
BLU RAY is the 132 minute Original U.S. Theatrical Cut FINALLY!!!!
]
On this Blu-ray for the first time ever we get the ORIGINAL 132 minute version of the film as was shown in U.S. theaters.
(Note: This is NOT the "Love Conquers All" Sheinberg/TV Edit.)
All the DVD releases before this have been the European Version/Director's Cut of the film. Even the original Universal DVD release that said Theatrical Cut on the box actually had the European Version/Director's Cut.
Unfortunately, Universal didn't really spend any time cleaning up the negative for Hi-Definition (there is very noticeable instances of dirt and debris on the negative) but it still beats the image quality found on the Universal SD DVD, and they do include an amazing DTS-HD Master 5.1. We'll just have to wait for the Criterion Blu-ray of Brazil for image perfection but serious fans of Brazil will want to get this Blu-ray edition just to have this version of the film as it was originally seen in theaters in 1985.
I actually find this 132-minute version is in many ways a superior cut of the film. Here are the differences in detail:
*In the 132-minute version you cut from Sam in bed with Jill, police sack goes
over head, then CUT TO Pull off police sack to reveal Sam in Torture
Chamber/Interrogation chair. This one cut is simply brilliant and very powerful.
In the Criterion Version you have the added scene of Mr. Helpman as Father
Christmas (completely out of charcater from the rest of the film) and the whole
interrogation scene of Sam hanging from the rack inside the police/mail pouch
which becomes narratively redundant and dilutes the impact of the final scene.
*To end it with cooling tower/interrogation room fade to clouds was a great Gilliam
wink of subversion and irreverence to the cliche Hollywood Ending. As opposed to
the European cut of just credits over cooling tower/interrogation room.
*The Samurai Scene is divided into 3 separate scenes in the 132-minute version
versus 1 LONG scene in the European cut. And you know what? Like most things,
it works better in 3's.
*The 132-minute version cuts straight to the Dinner Scene with Ida (his mother)
ordering numero deux, trois, etc. while the European version has the entrance to
the restaurant of going through the metal detector which really doesn't add
anything and is again a bit redundant when the bomb does eventually go off in
the restaurant. With the scene, you're signaling to the audience we are looking for a bomb, so we expect a bomb. Without the added scene, the bombing is unexpected and it actually shocks you so you're both horrified and laughing. The unexpectedness also works as it builds upon the bomb motif from the first explosion at the beginning of Brazil during the Ducts advert.
*And finally I just love that the 132-minute version opens on those clouds (outtakes
from The Never-Ending Story) then goes to the Central Services advert about
Ducts: "Are your ducts old-fashioned, out-of-date... " Now the Criterion version
also has the clouds opening (The Original European Cut didn't) but it's funny
because the Studio asked Gilliam to start off with the clouds for the US Cut and he
actually prefers it as quoted in his Director's commentary. Hilarious.
Little changes that add up to a tighter and overall, better film.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 12, 2011
★★★★★ 5
Great cast; well directed
Great movie; lives up to its reviews
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 25, 2026
★★★★★ 5
An ageless classic !
I hadn't re-watched this movie since the first time I saw it, when it came out in 1985. Back then, it had stricken the public with its fatalistic and pessimistic view of an overwhelming bureacracy, a suggested characteristic of fully-centralised power in a controlled society in the future.
Today, we can still admire some of its validity and premonitory qualities (and the sarcastic humour !). The dream-like scenes and symbols used in the film are also ageless.
Ever-growing computer systems govern today the relationships we hold with banks, government departments, utilities, phone companies, etc. Who hasn't experienced the frustration of dealing with their 'machines' and/or incompetent & robotic bureaucrats when problems or errors arise in their service? 'Brazil' will seem so familiar!
The omni-presence of screens in our lives, everywhere and incessantly offering us products or services, or used to control our movements and behaviour, is also a well-guessed futuristic scenario by Gilliam & co.
And what to say about the 'terrorism' and the 'permanent state of fear' portrayed back then? Sounds familiar today?
I recommend this movie to anyone interested in the threats of an unbridled, arrogant and all-powerful bureaucracy in human society.
The film can be a bit 'heavy' at times, though. Not everyone's cup of tea.
Watch it first, before sharing with friends or family.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 19, 2012