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Description
Mouse KIBRA 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. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 5000pg/mL). Then dilute to the following concentrations: 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312.5pg/mL, 156.25pg/mL, 78.125pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 5000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 2500pg/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 enzyme conjugate are sequentially added to microwells pre-coated with protein KIBRA (KIBRA) capture antibodies. 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 protein KIBRA (KIBRA) 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 KIBRA ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | WW and C2 domain-containing 1 (wwc1), also known as protein KIBRA or kidney and brain expressed protein (KIBRA), is encoded by the wwc1 gene. This gene encodes a cytoplasmic phosphoprotein that interacts with PRKC-zeta and dynein light chain-1. Alleles of this gene have been found to enhance memory in some individuals. Three transcript variants encoding different isoforms have been identified for this 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 | 78.12-5000pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.4 ★★★★★
Based on 19 reviews
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Product Reviews
★★★★★ 3
good to a point
Format: Paperback
basically what could help democrats win.all well and good,but that side has much of the same donors(drug companies,defense contractors,oil industry,etc.)as the republicans.THAT'S why they don't push back fundamentally.
one of my big problems with the author is his unapologetic.uneducated islamaphobia.he sounds like george bush when he mentions muslims actually.he fell for the propaganda.instead of drinking the koolaid of the cult,he should sip from the tea of informed tact.
i know right-wingers wear their stances/prejudices on their sleeves,but the problem with the liberal side is the smugness they can exude towards everyone else,when,let's face,they're no better.they went to college to deepen THEIR prejudices with a more expanded vocabulary.
otherwise,it's interesting from a psychological standpoint on how and what moves the masses.again,it's worth it to a point,just keep in mind that he's a bit of a meathead
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Reviewed in the United States on November 22, 2020
★★★★★ 4
The one-stars miss the point:
Format: Hardcover
Thomas J. Farrell and I may be two of a small handful who actually have read Aristotle's Rhetoric. There are good reasons for this. Aristotle's rhetoric is useful to know historically, and gives one the aroma of scholarship, yet only in the sense of one's being well-read but not particularly useful.
Westen's point is that Democrats are starving for useful rhetorical advice. Grounding ourselves in material some 2,300 years old is just not sufficient. cglambdin also missed the whole point, but more bluntly and therefore clearly.
I would paraphrase Westen's major point as being: as long as you go around thinking "reason, good/everything else, not so good," you lose. Not only do you lose, you DESERVE TO LOSE.
Why?
In a democracy, "nobody likes a smartass." The corollary to this is: "if you don't know the difference between being smart and being a smartass, you're probably the latter." Now to an ancient aristocrat like Aristotle, the distinction wouldn't have mattered. In the United States of America, it should matter to everyone aspiring to leadership.
We common folk expect our leaders to resonate with our values and life conditions. We don't care whether your blood runs a bit blue (as with the Kennedys) as long as you can be with us in spirit when you need to be. It's only polite.
In 1992 the smartass class had great fun with Bill Clinton's "I feel your pain" comment, but missed the point that Clinton resonated while President Bush the First's glance at his watch during the same town meeting debate ended the campaign then and there.
Drew Westen evokes what I considered state of the art in the communication field when I was in graduate school twenty-five years ago. Because he's a psychologist, and also not a smartass, I didn't expect him to bring up the theoretical language of people ranging from George Herbert Mead to Kenneth Burke. Rather, he demonstrates their insights! We get it! His work also fits well in the tradition of Walter Fisher's groundbreaking
.
Two things about Westen's book take off a star. Yes, he does meander. Also, his repetitive bashing of Bob Shrum comes off, at last, as an extended hard-sell advertisement for his own political consulting business. Perfection is elusive. Nevertheless, The Political Brain is doggone useful!
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Reviewed in the United States on July 27, 2007
★★★★★ 5
Disturbing but necessary read
Format: Paperback
Feels strange saying that I love a book that is as disturbing as this one is but I love that it's well-written and documented and it exposes some horrendous events in the history of the Americas as well as the world. Americans may well point fingers at the Nazis (and deservedly so) but it's a case of people in glass houses throwing stones. There is no question as to the repulsiveness and inhumanity of the genocide and mass murders perpetrated in Nazi Germany, Stalin's Soviet Union, Mao's China, the Khmer Rouge's Cambodia, and in a host of African nations, to name a few; but in our own not too distant past similar atrocities were perpetrated on the native populations of the Americas by the Spanish, Portuguese, British, and colonists/Americans. While the inhumanity elsewhere in the world is touched upon to show where the mindset of this barbarity likely originated, the focus is on the impact in the Americas -- North, South, and Central. The book contains graphic, disturbing descriptions of the cruelty done to the natives by men who have long been esteemed for their alleged contributions in history. Most notably Christopher Columbus. In my time in grade school, he and the many other conquistadors and explorers were portrayed and men of courage and integrity. This book paints a different picture of them as greedy, bloodthirsty, remorseless killers of peoples who they considered inhuman or subhuman. More troubling is Christianity's participation in these actions. Not to blame Christianity for initiating it but to indict it for condoning and even commending the events. Peaceful races of people minding their own business, living in communities well planned and constructed and advanced for their time, and who welcomed the interlopers were obliterated them. Much of the death and destruction was caused by the introduction of European diseases such as smallpox that killed tens of millions but a large part was also caused by actions of odious proportions. Entire civilizations such as the Aztec and the Inca and the Arawak whose artifacts have since been admired and sought after were wiped off the face of the earth. Tens of millions of people slaughtered. Men, women, children butchered. In numbers likely to exceed those of the aforementioned genocides combined. The purpose being to acquire their lands and their riches. Considered to be no more than animals needed to be exterminated. This is our history.
This book should be required reading for everyone. The graphic descriptions of the savagery should cause outrage not only for the acts themselves but for a cultures that has covered up their crimes for centuries. And the holocaust has not ended yet.
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Reviewed in the United States on October 23, 2018
★★★★★ 5
THE SINGLE FINEST AND MOST ESSENTIAL BOOK OF THE AGE
Format: Paperback
A masterpiece of scholarship and analysis.
This book is nothing less than the single most important work that you will ever read.
Our entire culture is built on Holocaust Denial while those most responsible for this abnesia drape themselves in the flag of holocaust memorialism but have little honesty in their true agenda. An agenda that allows in North America alone for there to be at least 50 Holocaust memorials, museums and monuments...
only problem is they are ALL about the Holocaust that happened in Europe and NOT about the colossal extermination that took place where they live. It is not only denial on the part of the nations of the Americas and Europe but those responsible for this Holocaust Denial in relation to Indian America insist on an image of being the world's caretakers of holocaust memory. What a bloody audacity.
Why do we let the Spanish off the hook so lightly? Why is there no demand for Spain to make its Mea Culpa? Why is there no AMERICAS HOLOCAUST memorial in Madrid, Washington, London and Ottawa ?
This brilliant book re-addresses the imbalance.
POST SCRIPT....
There is a reviewer further down who uses the monica of
"history buff" who rejects the value and integrity of this work. In fact he utterly insults Mr Stannard and his thesis.
So I thought I would check out his other reviews...oh boy!
One of the remarks he makes in a book claiming that Saddam was behind 9/11 goes "But it is very difficult to argue with the facts that were available to the agencies which pointed to a direct link between Saddam and Al Qaeda." This example of his world view is the mild end of it. So people consider the character of the self-described "history buff" who rejects Stannard's brilliant thesis on the Holocaust in the Americas.
The reviewer "history buff" has a world view that comes straight out of the 1950's HUAC committee (he associates all Left wing thought with the Soviet Union not knowing that the Bolshevik regime prohibited the platform of the revolution and that its first victims were in fact the most sincere and dedicated Left revolutionaries. Clearly he has never read the finest autobiography in the history of English language autobiography; Emma Goldman's LIVING MY LIFE volume 1 and volume 2. The latter volume includes a first hand account of the destruction NOT construction of socialism by Lenin and his cohorts ).
.
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Reviewed in the United States on June 5, 2006
★★★★★ 5
EXTINCTION
Format: Paperback
Normally when the word extinct is used it is in reference to animals but after reading this book this term can legitimately be used to describe what happened to the original citizens of America the American Indians. Christopher Columbus and his minions (I refuse to call them soldiers) savagely murdered and killed countless numbers of these Indians, a vivid example of this is given on page 83 " One favorite sport of the conquistadors was "dogging." Traveling as they did with packs of armored wolfhounds and mastiffs that were on a diet of human flesh and were trained to disembowel Indians, the Spanish used the dogs to terrorize slaves and to entertain the troops. An entire book Dogs of the Conquest, has been published recently, detailing the exploits of these animals as they accompanied their masters throughout the course of the Spanish depredations. "A properly fleshed dog," these authors say, "could pursue a 'savage' as zealously and effectively as a deer or a boar.... To many of the conquerors, the Indian was merely another savage animal, and the dogs were trained to pursue and rip apart their human quarry with the same zest as they felt when hunting wild beasts." And also on pages 83-84 "Just as the Spanish soldiers seem to have particularly enjoyed testing the sharpness of their yard-long rapier blades on the bodies of Indian children, so their dogs seemed to find the soft bodies of infants especially tasty, and thus the accounts of the invading conquistadors and the padres who traveled with them are filled with detailed descriptions of young Indian children routinely taken from their parents and fed to the hungry animals. Men who could take pleasure in this sort of thing had little trouble with less sensitive matters, such as the sacking and burning of entire cities and towns, and the destruction of books and tablets containing millennia of accumulated knowledge, wisdom, and religious belief."
After page 146 there's an illustrated unnumbered section titled Genocide the first nine pages of this section contain pictures of how the Spanish tortured and killed Indian women and children as stated on the second page of this section "[The Spaniards] took babies from their mothers' breasts, grabbing them by the feet and smashing their heads against rocks...They built a long gibbet, low enough for the toes to touch the ground and prevent strangling, and hanged thirteen [natives] at a time in honor of Christ Our Saviour and the twelve Apostles. Then, straw was wrapped around their torn bodies and they were burned alive."
Not to be outdone the good old American cavalry also engaged in its' fair share of savage murder and killing as can be seen from the bottom of page126 to the top of page 127 "They turned their guns, Hotchkiss guns, etc., upon the women who were in the lodges standing under a flag of truce, and of course as soon as they were fired upon they fled....There was a woman with an infant in her arms who was killed as she almost touched the flag of truce, and the women and children of course were strewn all along the circular village until they were dispatched. Right near the flag of truce a mother was shot down with her infant; the child not knowing that its mother was dead was still nursing, and that was especially a very sad sight. The women as they were fleeing with their babes were killed together, shot right through, and the women who were very heavy with child were also killed." I personally do not celebrate Columbus Day and Thanksgiving Day this book is ample proof and evidence as to why these two days should be set aside as a time for mourning not celebration, if you want your children to have a true understanding of American history then I strongly urge you to buy this book and have it as part of your home library.
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Reviewed in the United States on July 5, 2015