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Description
Mouse HSP90 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. Cell lysis solution: Gently wash adherent cells with pre-cooled PBS, then digest with trypsin, and collect the cells after centrifugation at 1000×g for 5 minutes; suspended cells can be directly collected by centrifugation. Wash the collected cells 3 times with pre-cooled PBS, add 150-200uL PBS for every 1×10^6 cells to resuspend (it is recommended to add protease inhibitors to PBS; if the content is very low, the PBS volume can be appropriately reduced) and break the cells by repeated freezing and thawing or ultrasound. Centrifuge the extract at 2-8℃, 1500×g for 10 minutes, and take the supernatant for detection. 5. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes, take the supernatant for detection, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 6. 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 90kDa Heat Shock Protein (HSP90) capture antibody. 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 90kDa Heat Shock Protein (HSP90) in the sample. The absorbance (OD) is measured at 450nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse 90kDa Heat Shock Protein | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Heat shock protein 90 (HSP90) acts as a molecular chaperone, regulating and maintaining the conformation and function of multiple proteins within cells, thereby helping cells grow normally under stressful conditions. Recent studies have shown that many oncogene proteins are targets of Hsp90. Therefore, inhibiting Hsp90 function could promote the degradation of these oncogene proteins, potentially aiding cancer treatment. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids |
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4.8 ★★★★★
Based on 1734 reviews
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Product Reviews
★★★★★ 5
Ben Solo’s fall is tragic
Format: Paperback
We don’t know much about Ben Solo’s past, but this comic explains a little bit more about why he is the way he is. It also explains that he didn’t do all the things he himself implies in TLJ. It’s a story of a young man who is lost because those he is supposed to trust betray him, which is exactly what Snoke/Palpatine want. It shows he didn’t really have a choice in the path he took and it makes his redemption at the end of the saga powerful, even if the last movie itself destroys the myth of Star Wars. I love knowing that Rey and Ben have always had a connection and I hope that is something explored more deeply in Episode X when they bring Ben back.
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Reviewed in the United States on July 30, 2020
★★★★★ 5
Answers a lot of questions
Format: Kindle
I enjoyed this story. This fills in a lot of gaps left open from the sequel trilogy, and the artwork was pretty cool. Excellent coloring and layout.
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Reviewed in the United States on August 26, 2023
★★★★★ 5
Revenge for a fallen emperor, redemption for a fallen warrior
Format: Kindle
The crimson empire saga is the story of kir kanos, last of emperor Palpatine's crimson clad imperial guardsmen. It is a story of revenge and redemption, as over the course of the trilogy Kanos grows and changes in ways he'd never have expected. I did especially like his changing relationship with Mirith Sinn, and how her arc is a fitting mirror to his own. I also liked the villain of the first series. Jax, and I wish he had stuck around for the rest of the trilogy. The rest of the villains throughout the trilogy are far less memorable than jax, but I suppose that's the problem when you make sequel to whats unseemly a self contained story.
Of the three series contained here, the first series is by far the best. It's quite self contained, and any knowledge of the universe at the time is explained quite well. It's also the simplest, Kanos wants Jax dead, and Jax wants Kanos dead. The second series is abit of a mess, with an overly complicated plot and a good deal of unseemly wasted page count on the random side characters that don't advance anything. In particular I'm thinking of the hutt character, as well as the mystery hooded figure who doesn't actually come into play at any point in the saga. Perhaps he comes into play in some later star wars story, but in this it reeks of a dropped plot line. The third and final series is a worthy sequel to the first although the scenes with Han solo also seem unnecessary. I did like the ending of the saga, even though I would have referred a bit of a more definitive ending in terms of certain character relationships.
The art of the saga get progressively better with each series. In the first series it's still reasonable good, but some of the faces and body proportions are bit weak. By the third it's really good if standard comic art. The fight scenes are well done throughout, and the ship battles are a great to see.
In conclusion, I would recommend this to anyone who likes a good revenge story. Don't worry about the context of this in the greater star wars saga, the most pertinent information is explained well enough, in my opinion. As long you have seen the star wars movies and remember who the emperor is, your good.
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Reviewed in the United States on September 22, 2017
★★★★★ 5
A must read
Format: Kindle
This series is a must read if you love Star Wars post Endor. The galaxy is still in a struggle to see who is on top. With many a individual falling through the cracks in galactic society.
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Reviewed in the United States on October 23, 2025
★★★★★ 4
One man's quest for vengeance
Format: Hardcover
"A long time ago in a galaxy far, far away..."
It is the time of a falling empire, a time of betrayal and vengeance. This is the story of the last of the mysterious Imperial Guards of the Emperor Palpatine. It is the story of his quest to seek justice for the galactic ruler he served, and for his fellow comrades in arms, who gave their lives in that service.
"Star Wars: The Crimson Empire Saga," tells the tale of Kir Kanos, the sole survivor of the scarlet clothed guardians of the Emperor, first shown in "Star Wars, Episode VI: Return of the Jedi." Over more than a decade of Dark Horse Comics publishing history, starting in 1997 to its long awaited conclusion in 2012, "Crimson Empire" reveals the history of these elite soldiers, "the best of the best," trained for one purpose: to serve the Emperor of "the first Galactic Empire." This 504 paged hardcover book collects the following stories:
Star Wars: Crimson Empire #1-6 (1997)
Star Wars: The Bounty Hunters (Kenix Kil) (1999)
Star Wars: Crimson Empire II: Council of Blood #1-6 (1999)
Star Wars: "Hard Currency" (Dark Horse Extra #21-24) (2000)
Star Wars: Crimson Empire III: "The Third Time Pays for All" (Dark Horse Presents #1) (2011)
(Note: The above is also the opening passage to Crimson Empire III #1)
Star Wars: Crimson Empire III: Empire Lost #1-6 (2011)
Star Wars Handbook Volume 2: Crimson Empire (1999)
"The Crimson Empire Saga" begins with an ending; the final defeat of the Emperor Palpatine by his long-time foes, the Rebel Alliance, led by the legendary siblings, Luke Skywalker and Leia Organa Solo. But unbeknownst to those fighting to free the galaxy from Palpatine's tyranny, treachery also played a role in the Emperor's downfall. In a stunning turn of events it is revealed that one of those thought to be most loyal to Palpatine, a member of the Imperial Guard itself, betrayed his master in an attempt to seize power for himself. To insure the success of his treason, Carnor Jax also ordered the assassination of all the surviving Imperial Guard. But in his arrogance, the traitor underestimated the skill of the warriors he had trained with. Though faced with the overwhelming force of an army of stormtroopers sent to eliminate them on their training world Yinchorr, one Imperial Guard survived the massacre. Kir Kanos, the last of Emperor Palpatine's Imperial Guard lived on, to seek vengeance on Carnor Jax, and all those who had betrayed his Emperor.
The core "Crimson Empire" trilogy is the creation of writers Mike Richardson and Randy Stradley, and artist Paul Gulacy, and they have produced an exciting adventure of Star Wars action, intrigue and emotion. The first "Crimson Empire" 6 issue comic mini-series was published in 1997, in the early years of what came to be called the "Star Wars Expanded Universe." The "EU" is an ongoing showcase for the continuing "adventures of Luke Skywalker" in novels, comics, computer games and more, as well as for other characters and worlds from the movie and now television saga envisioned by George Lucas. But the Expanded Universe also highlights original ideas based on characters and situations created by Mr. Lucas. "Crimson Empire" represents a prime example of talented imaginations thinking, 'what if we told a story about the red cloaked guards seen in "Return of the Jedi?"'
In Kir Kanos, Richardson, Stradley, and Gulacy have created a classic Star Wars anti-hero/hero. Along with such Expanded Universe creations as Grand Admiral Thrawn, Mara Jade, Quinlan Vos, and Asajj Ventress, he is one of the most intriguing characters in the Star Wars Universe. Kanos is a highly trained warrior with a steadfast moral code, whose quest for vengeance anchors "The Crimson Empire Saga." It is this focus on the lead character's ultimate goal that is one of the achievements of the series. This dramatic strength, when adhered to, allows the comic creators to truly delve into their original characters and concepts without having to include or depend on the more famous Star Wars heroes and villains, although some appear in supporting roles. It is a testament to the makers' storytelling abilities that as the reader follows this tale of Kir Kanos' road to revenge, one becomes sympathetic to the character's cause to avenge the death of one of the most evil men who ever lived in any galaxy!
A special highlight of "Crimson Empire" is the artistry of Paul Gulacy. Mr. Gulacy has a distinctive style in the sequential art medium; a clean precise vision that combines traditional comic book panel progression with simulated photographic technique. For example, Mr. Gulacy rarely uses motion lines in his comic art, except to depict the movement of a lightsaber, or in the case of Kir Kanos' preferred weapon, the force pike, a doubled-bladed long staff. This style gives Mr. Gulacy's work a heightened sense of realism. In the first "Crimson Empire" mini-series, Mr. Gulacy is inked by P. Craig Russell. In "Crimson Empire II: Council of Blood," he is embellished by Randy Emberlin. "Crimson Empire III: Empire Lost" saves the best for last as Mr. Gulacy produced full pencil and ink artwork for the third mini-series. He was always his best inker.
This "Crimson Empire" collection also offers the reader the opportunity to see the evolution and advancement of comic coloring from 1997 through to 2012, the year of this special compendium's release. Dave Stewart created the color art for Mr. Gulacy's pencil work on the first two installments of the prime comic trilogy and the results display a marked improvement over the flat colors long produced in comics of the past. But with "Crimson Empire III," the most recent part of the saga, the color rendering of Michael Bartolo shows an even more nuanced use of tone and hue over Mr. Gulacy's art. Thanks in no small part to the computer age, comic coloring has come a long way from the days of newsprint.
Another highlight of "The Crimson Empire Saga" is the inclusion of two addition stories within the thick hardcover. "How the Mighty have Fallen" is a tale written by Randy Stradley, and illustrated by Javier Saltares and Christopher Ivy. Published in 1999 as part of the Star Wars: The Bounty Hunters comic series, it relates how Kir Kanos created the secret bounty hunter identity Kenix Kil,to better hide from members of the fallen Galactic Empire that were hunting for him across the stars.
The second adventure, the short story, "Hard Currency," was published in Dark Horse Extra, a newspaper-like fanzine released from 1998-2002, and features Kenix Kil teaching an enemy the cost of betrayal. It was scripted by Mr. Stradley, and drawn by Isaac Buckminster Owens.
There is a great deal to enjoy in this "Crimson Empire" collection but it is not without flaws. While the premiere "Crimson Empire" mini-series succeeds in being an enthralling adventure on every level, "Crimson Empire II: Council of Blood" sometimes suffers from a story that concentrates too much on subterfuge and political power-plays; so much so that the series' starring character, Kir Kanos, seems rather to be a supporting player in his own story. The long-awaited third part in the "Crimson Empire" trilogy, "Empire Lost," improves on the faults of its predecessor but still has a sometimes convoluted plotline.
Another flaw in the multi-part saga is the character arc of Mirith Sinn, the brave and beautiful Rebel Alliance commander who plays a major part in most of Kir Kanos' exploits. The series writers, Mr. Richardson and Mr. Stradley, showcase the evolution of the relationship between these two driven individuals, a relationship that has an undeniable chemistry. Mirith and Kir Kanos share a bond, a bond that is complicated by violent loss. But the complexities of their relationship are sometimes handled in conflicting ways from story to story, resulting in confusion for the reader in understanding the otherwise strong character of Mirith Sinn.
Still another flaw in this collection is the editorial decision not to include all the striking covers painted by artist Dave Dorman for the 18 individual comics in the "Crimson Empire" trilogy of mini-series. While it is gratifying to see 6 of Mr. Dorman's contributions to the saga, notably his special wrap-around cover painting to the first "Crimson Empire" trade paperback reprint volume, as well as the "Crimson Empire Handbook" cover, this reviewer regrets the omission of the artist's 13 other paintings. Nor was the sole "variant" cover to "Crimson Empire III" number 1, drawn by Mr. Gulacy included, and it was the only cover he created for the entire series. It is doubly disappointing when the collection showcases Mr. Dorman's cover to that same comic issue twice in the book!
This reviewer would have also been appreciative of a written introduction or history of the creation of the "Crimson Empire" series; and a feature that explained the long wait of 13 years before the concluding chapter in the trilogy was finally released for fans of the saga to enjoy. These editorial choices could have then given this hardcover collection the more appropriate title, "The Complete Crimson Empire Saga."
Despite the above shortcomings, this is a very special Star Wars comic collection. From one Star Wars fan to any who read this review, I highly recommend you put on some of John Williams' immortal Star Wars music, sit back, open this hardcover and let yourself be taken away to that "galaxy far, far away" we love so much!
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Reviewed in the United States on September 18, 2012