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Description
Rat IL-7 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: 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: 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. 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. 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. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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 200 pg/mL). Then dilute to the following concentrations: 200 pg/mL, 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 200pg/mL standard working solution into the first EP tube and mix thoroughly to make a 100pg/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 conjugate are sequentially added to microwells pre-coated with Interleukin 7 (IL-7) capture antibody. 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 Interleukin 7 (IL-7) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Interleukin 7 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-7 (IL-7) is a hematopoietic growth factor secreted by stromal cells in the bone marrow and thymus. It is also produced by keratinocytes, dendritic cells, hepatocytes, neurons, and epithelial cells, but not by normal lymphocytes. A study has also demonstrated how autocrine secretion of the IL-7 cytokine by T-cell acute lymphoblastic leukemia (T-ALL) contributes to the oncogenic development of T-ALL and provides new insights into the dissemination of T-ALL. IL-7 stimulates the differentiation of multipotent hematopoietic stem cells into lymphoid progenitor cells (in contrast to myeloid progenitor cells, whose differentiation is stimulated by IL-3). It also stimulates the proliferation of all cells of the lymphoid lineage (B cells, T cells, and NK cells). It is crucial for proliferation at certain stages of B cell maturation, T and NK cell survival, development, and 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. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 3.12-200 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.5 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 3
Not a bad start
Format: Kindle
3 stars
Thank you Netgalley and Briar Boleyn for the ARC!
A camelot/king Arthur retelling with fae. I was hooked by the idea of this book immediately and was eager to jump into this world.
• slow burn
• enemies to lovers
• who did this to you
Morgan Pendragon watched her mother die by her father's hand when she was just eight years old, hiding under the bed. Morgan is believed to have the tainted blood of the fae in her veins and is cast aside so that her fathers illegitimate son, Arthur, can become the king. She's seen his cruel treatment of the fae firsthand, so when he sends her on a journey to find a fae weapon she seizes the opportunity to do more with her life. Along the way, she finds more than she could have imagined.
I don't know a whole lot about King Arthur and Camelot but I had a lot of fun with this story! The plot has some similar tropes to popular romantasy books (From blood and ash) but there's enough originality here that it doesn't feel like I'm reading a copy. I liked how the fae were different in appearance than what is typical in most fantasy books I've read. In this book they have blue hair, violet skin and a wide range of other characteristics. I thought that the world building was easy to follow and I could easily immerse myself into this world. After reading the blurb I kept wondering when she was going to go on the journey to find Excalibur and it doesn't happen until around the 45% mark.
The story is a bit slow at times but starts to pick up once they begin their journey to find Excalibur. The John Wick style Inn was a fun concept that I enjoyed reading about.
There are a lot of similarities to this and FBAA and I would have liked to have it be a little more different, but I'm hoping book two will have the story turn into something of its own. Overall I enjoyed reading this story and I'm looking forward to reading book two especially after that ending.
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Reviewed in the United States on May 27, 2023
★★★★★ 5
Epic cliffhanger
Format: Kindle
All I can say about this book is WOW!
Absolutely loved the characters in this book. Raven is such a bad ass chick.
Brax
Creed
Zane
Eldon
They’re all such good characters my favorite is Brax even with his brooding personality at first.
This book is a PNR with such great chemistry between the characters right away. I love the snap backs between them all. There is a bit of mystery thrown into the story as well. We don’t get to know what everyone’s gifts are right away an I like that a bit it helps us get to know the characters first in my opinion.
Eldon is the one who falls first then the others Raven is sorta stand off ish mostly because she has no idea as to what life is like at Silvercrest. She was brought up in Shadowmoor an we find out why towards the end. Her brother is a pain he’s so full of himself I have a feeling he an the dad have a sinister plan for some reason.
The connection between the guys an Raven is a strong one. Eldon has his visions that he this has to do with Raven but you get a glance at them as well.
I don’t like Finn an Genie you’ll see when you read.
I love how Raven instantly makes friends with Leila regardless of what others think. I hope we get more of Raven and her familiar the cliffhanger was a mind blown thing. I’m excited to see where that goes I’m not typically a PNR person but this story just drew me in an I’m glad it’s so good it’s not a dark romance as I tend to go towards but I love it it’s a breath of freshness an I think a lot of you would definitely enjoy this series remember it’s a reverse harem so you don’t have to pick a favorite guy 😏
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Reviewed in the United States on October 2, 2023
★★★★★ 4
I’m excited! this was Fun!
Format: Kindle
I really liked this! This is a new author for me. I was looking for something with Fantasy and magic and I think I really scored a good one! A lot of this book leaves you guessing and trying to fill in the blanks with Raven about what’s being kept from her and why’s she’s had to live the life she’s had. I was really confused by her family, it’s definitely disfunctional. Her brother wants to kill her, her father is a major a22 hat, her mom looks like she needs major help to escape from her husband. They really threw me off. I knew it was going to be something big and the clues are dead give away. Even though it leads you to outcome it’s still a fun read, lots of banter and HOTTNESS going on! I loved her guys, they came on really strong and it just works. I’m looking forward to the next book. The only reason I didn’t give it 5 stars is how predictable it is any sometimes raven was a bit annoying. It was enjoyable but I did put it down a few times and come back later. It has my interest.
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Reviewed in the United States on February 15, 2024
★★★★★ 5
Pink Raven…
Format: Kindle
I am convinced that KC Kean can write anything and I will absolutely love it. From dark romance to fantasy she’s taken me on a ride of ups, downs, twists, turns and some pretty brutal cliffhangers. This book didn’t disappoint in bringing it all and all I can say is give me more.
Raven’s journey to Silvercrest Academy brought with it some rough patches in her life. She spent 14 years in a place of darkness fighting everyday to survive. Only for her father to bring her back to a deceiving light.
When her father returns for her, she’s surprised with his decision to send her to Silvercrest Academy especially since she is a void. She has no magic or atleast that’s what she thought. When she arrives she sees her brother for the first time in 14 years and he hasn’t changed, he’s still the angry spiteful little boy that she knew back then.
She’s placed in a house with four of hottest guys she’s ever seen. Eldon, Brax, Creed and Zane are first years and her new roommates. Their relationships grow over time and we get to see some of their magic over trials that are meant to break them.
I really enjoyed this first book. Raven is tough on the outside and for good reason. We sense a different side to her once she feels a connection with the one’s she cares about.
We do get left on a slightly soft cliffhanger and I can’t wait for the next book to see what happens next.
Fantasy setting, steamy bits and an unknown dark force that has started to show its intentions. Yes! Give me more!
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Reviewed in the United States on October 4, 2023
★★★★★ 5
falling in love with this book 🖤
Format: Kindle
This book captured me from the very beginning. Raven is an absolute baddie. She grows up in one of the roughest places and then gets tossed into an academy that is made to break the students who attend.
When she arrives she finds herself in a house with four roommates Zane, Eldon, Creed and Brax by accident.
The guys are protective from the very beginning and I love that. Raven doesn’t take any crap from anyone and seeing her have that support that she hasn’t really ever had is really sweet🖤
Then there’s her brother Sebastian 🙄 who for some reason can’t stand Raven even though he hasn’t been around her since she was 6. He is an absolute twatwaffle.
Then the gauntlet is brought out to play and for some reason it’s being done differently than it is usually done. Everyone has to participate in it and Raven is without magic to help her escape the gauntlet with her life. Getting to see each of the guys magic and the affinities that they possess is so freaking exciting. And then Raven meeting Ari 😍😍😍 Though I do feel like that last outpost visit to shadowmoore was a set up to get her to use her magic that they think she has. Ugh this book kept me pulled in to the very last word.
7/5⭐️’s
2.5/5🌶️’s
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 4, 2023