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Description
Human RBMS1 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. |
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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 a capture antibody against RNA-binding motif, single-stranded-interacting protein 1 (RBMS1). 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 RNA-binding motif, single-stranded-interacting protein 1 (RBMS1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human RNA-binding motif, single-stranded-interacting protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | RNA-binding motif-single-stranded interacting protein 1 is a protein encoded by the RBMS1 gene. This gene encodes a member of a small family of proteins that bind to single-stranded DNA/RNA. These proteins are characterized by the presence of two sets of ribonucleoprotein consensus sequences (RNP-CSs), containing conserved motifs, namely RNP1 and RNP2, originally described in RNA-binding proteins and required for DNA binding. These proteins have been implicated in diverse functions such as DNA replication, gene transcription, cell cycle progression, and apoptosis. Multiple transcript variants, resulting from alternative splicing and encoding different isoforms, have been described. A pseudogene has been identified at this locus on chromosome 12. RBMS1 has been shown to interact with the polymerase (DNA guide), α1. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.7 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
Great investment! It'll save your floors and help train your pup by giving him/her a specific area to pee/potty in the house!!
Color: Blue, Size: 17" x 23", Color: Blue, Size: 17" x 23"
Though my pup sometimes like to pee right at the edge of this puppy pad holder, when he does, he usually just hits the lip of holder. He's a beagle/dachshund mix, and will probably end up around 20lb - he's around 15-ishlb now - so not a tiny dog, but is still a small dog. And with him being "not" tiny/toy, and him being a male, he tends to not aim so well sometimes when he uses it. This made using the PP holder a learning curve for me.
I started out using the regular size pads in it
, which fit amazing well; however, with Morty's - ahem! - mishaps with aiming, I tried doubling-up on the training pads - would put two of the reg size pads on the holder - and that left just enough pad "hanging" outside of the holder to meet Morty's mishaps in aiming. I was even able to still fasten the holder through the pad, which is awesome!
However, using double to pads to cure Mort's lack of ability to aim for the center every time was costly, so I decided to shop around to see if I could find a solution to our aiming whoes. That's when I stumbled upon these pads at Walmart for just under $10 before tax, but they are available to purchase here at Amazon,
& it occurred to me that perhaps one XL pad would give Morty that extra allowance he needed, and it could be more affordable to me - it worked! .
These particular pads have been working out wonderfully because I continue to use just one pad and still get the overage protection - I'm pretty sure Morty intentionally pees on the edge sometimes - but since they're quite absorbent, it's very rare that he misses so much that he pees "through the pad." [The time he pees through the pad are the times he literally barely even hits the holder/pad at all because like I said, I'm pretty sure he aims for the edge on purpose most of the time.]
Since Morty is a beagle/dachshund mix, he is a scent hound, which means that I can never just "let him outside" - I always walk him outside with a leash - because scent hounds follow their noses [pretty sure his nose is more Beagle than Dachshund lol]. And with it being extremely hot out now, I allow him to pee indoors at least 50% of the time; I am absolutely thrilled that he has taken to his wee-wee pad like a pro.
All and all I'm very satisfied with this purchase because it has allowed us to have a clearly defined, and mostly contained space within out home for our pup Morty, to go potty in the house, and without ruining our hard floors. Moreover, since the holder itself is plastic, cleaning it up is a breeze! I just spray a bit of Clorox Clean-up on the holder, wipe it and voilà - odor-free home!
As an aside; the holder mentions being skid proof, and though I don't doubt the validity in that claim since the feet, which are on the four corners, do have rubber on the bottom, Morty is still a 7-month-old puppy, and sometimes, he can be a bit ornery and play with his pee-pad - which is always fastened inside the holder. And that subsequently allows him to move the holder from it's original spot because he "pulls" it since he's trying to "pull" the pee-pad off. Also, when my daughter and her friends play fetch with him, sometimes they'll throw his tow toward the same area that his holder sits. When that happens, he tends to run across the pad/pad holder and cause it to move. I'm attaching a video to illustrate what I mean. On this particular day Morty was playing by himself with an empty two liter bottle - he loves bottles! On this occasion, his pad/holder is not in it's normal spot - he's actually moved it into the kitchen. Toward the end you can here him baying out the door at someone/thing. I believe it may have been the neighbor lol :)
But since the times that Morty is able to move the holder are during times in which his doing it intentionally, I still wouldn't dock it for "not" being skid proof because it clearly is. I just wanted to point this out in case you furry family member is anything like mine, and is full of energy - I don't think there much of anything in my house Morty can move when he has high-energy like that. Also in the video, you can see how the XL pads fit onto the regular size holder, and the extra overage it provides - should you have a dog like Morty who needs a little extra overhang because of his aiming woes. I've also included photos of my pup, Morty. He'd climbed up on the chair for no reason really, but it's another example of his high-energy :)
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Reviewed in the United States on July 22, 2017
★★★★★ 5
Does what it says
Color: Dark Gray, Size: 22" x 22"
I was using just a puppy pee pad on the floor, but when my puppy started to become rambunctious....she'd quickly grab it and play with it...shred it and try to eat it. I also noticed that she tended to only want to use the edges of the pee pad majority of the time....so I typically ended up with it on the pee pad and then a puddle next to it. I had to do something. On Amazon I go like always and I found this. Didn't even know this existed. Works beautifully. I've had this for around 2 months now. Fits standard pee pads perfectly. It cleans easily. It doesn't hold in any odor. This does not completely keep my puppy from peeing on the floor. This is not the products fault, she aims towards the edges for whatever reason. If anything, it's helped keep more of it in. The raised edges do seem to encourage her to stay inside of it. We rarely have "accidents" now and no more shredding pee pads. See below.
*On another note- she has scratched at the pee pads, like she's digging in the dirt for gold as well as chewed on the plastic a fews times. I was worried I'd have to find another solution. She gets redirected with her chew toys and gets a treat every time she uses it correctly (no chewing or digging). At this time it doesn't seem like it will be an issue that I need to address. She hasn't done it since. So if your dog is determined this will NOT stop them. This product is not indestructible, please know this! It can be chewed on and if determined they can still get to the pee pad. I don't understand why some people rate it poorly when the product is doing what it's "intended" to do. You just have to know your situation and find the best solution you can live with. A dog is going to be a dog. Please address why they are doing what they're doing to begin with. I realized that my pup (*puppies will chew) wasn't getting enough mental stimulation, she's an Aussie. She had physical but not enough mental stimulation. Combine that with making sure we only encourage the wanted behavior and simply dismiss the unwanted behavior. I'm not saying I'm a pro, far from it... nor is everyone's situation the same....I'm just saying don't expect a product to do your job, expect what it says it's intended use is for. :)
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Reviewed in the United States on August 27, 2024
★★★★★ 1
NO GO!
Color: Dark Gray, Size: 22" x 22"
I was really hoping this would work, but it did not. The minute I put the tray down with puppy pad secured, my puppy managed to chew and dig at the edges ripping the pad away from the side of the tray and making a huge mess! I am not returning it bc this company makes you take it to UPS which is not convenient for me, so maybe I will donate it. If your puppy is massive chewer this may not be the thing for you.
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Reviewed in the United States on May 2, 2026
★★★★★ 3
Great idea.
Color: Dark Gray, Size: 22" x 22"
Great concept! Unfortunately, the plastic area underneath the pad area isn’t very thick and flexes. When my dog used it the first time the plastic flexing up and down under her feet frightened her a little bit. I placed a rubber mat between the pee pad and the flimsy plastic and that stopped the plastic from flexing but since it scared my dog from the first use she won’t go back on it.
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Reviewed in the United States on March 17, 2026
★★★★★ 5
PERFECT PEE PAD HOLDER: AFFORDABLE, EASY TO CLEAN & CHANGE THE PEE PADS, HOLDS PADS IN PLACE
Color: Navy, Size: 22" x 22"
I was THRILLED when we finally received it! I had been everywhere on Amazon & some other sites. I almost bought one that had a small pole since my newly adopted Chihuahua actually lifts his leg, unlike my previous male Chihuahua. But I kept running into a lot of holders that had a plastic "grill" to pee through the pad underneath. It didn't seem like it would be easy to clean. Plus, they were VERY expensive. However, I somehow came across this one. Once I saw the picture, it caught my attention. I watched the video & read more about. The only problem I remember is the video showed one that's foldable & easy to take anywhere with you. But that wasn't the case when we received it. However, I was still very happy with it. The only thing I wasn't crazy about was having to change pee pads because of the size. I changed to Earth Rated because that's what was recommended for this holder. My Chihuahua is around 5 years old & was "housebroken" before I adopted him from a shelter early last year. We live in an apartment on the first floor. But can't always run out the door when he has to go potty. He can wait, but I simply can't drop everything I'm doing to take him outside & I don't want to keep him waiting. He wouldn't really use it at first. But he finally got the hang of it. It's very convenient for him. I bought it primarily because I can't always go outside. Yet, it's perfect when it's raining & neither of you can't go outside, it's great to have when it's not safe to go outside in this area. I bought this around Black Friday week & got it for half its regular price! I'm thinking of getting another one to give him more space (sometimes he misses & accidentally sims for the floor). I've been cleaning it every time the pad needs to be changed. I use Biokleen Bac-Out for Pet Stains & Odors - Lavender Lime scent, which works great, but it's too expensive on Amazon. Also Earth Rated pads don't seem to work well with minimizing odors. I also find it a bit difficult to see if my dog peed because the bottom color is green. But they fit well & have adhesive corners. My dog never chews on the pads. The only problems he has is that he likes to push & pull the pad like dogs do to the grass outside when they finish peeing or pooping. Also, I don't know if the holder we have has non-slip small parts on the corners underneath. So since the floor is laminate & maybe has gotten more slippery from cleaning with the Enzymatic spray cleaner I use, the holder has been slippery lately. In the meantime, I'll try out the 100 pee pad pack sold by Iris, the maker of the holder. The previous pads don't have pads in the right size. I'm referring to Hartz 24 Hour Miracle Pee Pad in Lavender. They're white, but very absorbent & the best part: they help neutralize the odors.
I absolutely recommend this pee pad holder! We got the Navy, 22" x 22", which was on a HUGE sale for HALF ITS PRICE! It's too bad I didn't buy two! It's easy to take apart (the top from bottom) to clean. The handles that latch down & the design do a good job of keeping the pads in place. If only I can find pads like Hartz, I wouldn't have to change the pads so much lately. Although I LOVE the Enzymatic cleaner Biokleen Bac - Out for Pet Stain & Odors in Lavender Lime, I'm getting tired from cleaning. But hopefully when it gets warmer here in Florida, it'll get easier again. My dog is really wonderful! I bought this NOT to be lazy, but to give him a happier, more comfortable life. He seems to be happy to have the convenience of the pee pads & enjoys watching me clean!
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Reviewed in the United States on January 18, 2026
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