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Description
Human POMC 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 specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 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 proopiomelanocortin (POMC) 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 proopiomelanocortin (POMC) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Proopiomelanocortin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Proopiomelanocortin, also known as proopiomelanocortin (POMC), is a 241-amino acid precursor polypeptide. POMC is synthesized in the anterior pituitary gland from the 267-amino acid polypeptide precursor, pro-promelanocortin (pre-POMC). A 26-amino acid signal peptide sequence is removed during translation. POMC is cleaved (splitting) to produce a variety of peptide hormones. Each of these peptides is packaged in large dense-core vesicles and released from the cell by extravasation in response to appropriate stimuli. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.1 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
I fully recommend it!
Format: Paperback
The book was great! It was full of color and my 9 year old daughter loved it!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 13, 2025
★★★★★ 5
Great book!
Format: Paperback
Primer is a Great book! Highly recommend!
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Reviewed in the United States on May 6, 2025
★★★★★ 5
Great book
Format: Paperback
My 4th grade daughter’s favorite book.
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Reviewed in the United States on April 15, 2025
★★★★★ 5
Great for reluctant readers
Format: Paperback
This book is great for reluctant readers. I got this for my niece and her mother asked if I knew of any other graphic novels like this one because of how much my niece loved reading it. I ended up reading it and the story is very enjoyable and inspiring. The art is exceptional. I was very happy to find that there are more in the series. I bought both the first and second ones for my step daughter and other nieces this Christmas. Highly recommend!
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Reviewed in the United States on January 10, 2026
★★★★★ 5
Foster Care! Magic Paint! Superheroes! OH MY!
Format: Kindle
This was a great read. I loved everything about it. The artwork is vivid. The main character’s personality is spot-on. The humor was great.
Ashley is a girl in a world where she is herself and nobody else. At least, that’s what she thinks. Really, she’s a girl stuck in foster care because her dad’s in jail. She has a carefree attitude on the outside, but on the inside she’s really tender-hearted. Then one day a new family shows up, attempting foster care with Ashley. She’s living pretty nicely there and she’s made a friend named Luke. Then one day her foster mom comes home acting kind of strange. Later, Ashley decides to snoop into what’s in that mysterious suitcase her foster mom brought in and hid in a closet. She and Luke find paint. Lots of tubes of paint. Ashley puts them on her skin, because she “likes the texture.” This is where I think it’s waaaaay too obvious that what she’s doing has to be specifically made like that for the storyline. It’s okay though, they do an okay job of hiding it. Anyway. These paints are magic paints that give the person who wears them superpowers! So of course Ashley has to go and use them and be a superhero she calls ‘Primer’. But her foster mom’s job wants those paints she brought home back. So they send their roughest, toughest soldier to retrieve them. Ashley, of course, has a fight with her foster mom about it, and Ashley decides to run away, taking the paints with her.
Then obviously the soldier dude shows up, with a bunch of robots. There it just turns into your normal superhero fight scene, but then Ashley loses and the paints are taken except the teleportation one. The soldier, by the way, is named Strack. So then Ashley’s like, “Oh no, I’ll neeever be a hero” even though obviously she will, this is a superhero story. Suddenly her phone is ringing. It’s her foster dad and mom. She picks up their video call and it’s STRACK! He’s adult-napped her foster parents, of course. She debates going to fight Strack, or to just leave it. She goes with leave it until she looks up and sees a painting she made and this suddenly gives her confidence, for reasons unknown.
So then there’s another big fight scene with Strack, but Ashley is overconfident like she knows she can’t die, it’s a book and that would be devastating for little ones reading it. Anyway, she wins and frees her parents and they all live happily ever after.
So, this story ends in a cliffhanger that’s not a very good one. It’s just Ashley’s REAL dad seeing her on TV from when she went out and was a superhero the first time, and he’s like, “You’re not Primer, every father knows his daughter’s eyes, ASHLEY. See you soon.” So if I was hanging from a cliff here, I would be attached to it with a safety cable and I would be laying on the top of the cliff, with only my foot hanging off. It’s not much of a cliffhanger.
This was a great book about a female superhero. Oh, and another thing I forgot to mention, there is a page you should skip if you are reading to a child under seven. Page…. Let’s see here… oh yes. Page seventy-seven. It involves a gun and likely shooting afterwards, but it isn’t shown. I am a very sensitive person, and even I, an almost-teen was kind of rustled by it. Anyways, great story, lovely artwork, good book.
I’m rounding up from 4.5 stars.
-written by a tween
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Reviewed in the United States on February 26, 2022