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Description
Mouse apo-A1 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 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. 5. 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. 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 1000 ng/mL). Then dilute to the following concentrations: 1000 ng/mL, 500 ng/mL, 250 ng/mL, 125 ng/mL, 62.5 ng/mL, 31.25 ng/mL, 15.625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 1000ng/mL standard working solution into the first EP tube and mix thoroughly to make a 500ng/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 capture antibody against apolipoprotein A1 (apo-A1). 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 apolipoprotein A1 (apo-A1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Apolipoprotein A1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Apolipoprotein A1 is a protein encoded by the APOA1 gene. As a major component of high-density lipoprotein (HDL) particles, it plays a specific role in lipid metabolism. The APOA1 gene is located on chromosome 11, specifically at location 11q23-q24, and comprises four exons. APOA1 encodes a 28.1 kDa protein composed of 243 amino acids, with 21 peptides observed by mass spectrometry. Apolipoprotein A1 is the major protein component of HDL particles in plasma. Chylomicrons secreted from intestinal cells also contain apolipoprotein A1, but it is rapidly transferred to HDL in the blood. As a component of HDL particles, this protein accepts fat from intracellular compartments (including macrophages within arterial walls, which are overloaded with fat from oxidized LDL particles), allowing fat molecules to be shed elsewhere, either back into LDL particles or excreted in the liver. | |||||||||||||||||||||||||||||||||
| 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 | 15.62-1000ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.4 ★★★★★
Based on 110 reviews
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Product Reviews
★★★★★ 4
3 considerations for IQ BALL. FUN, DURABILITY & COST to TIME VALUE
Size: 4 INCH, Pattern Name: IQ TREAT BALL 4", Size: 4 INCH, Pattern Name: IQ TREAT BALL 4"
3 Points to consider buying IQ BALL : FUN, SAFETY & COST.
IQ BALL out of the well packed box: It is a good toy depending on your dogs' SIZES, TOY DESTRUCTION QUOTIENT & Cost to PlayTime ratio
1) FUN--Great fun for playful dogs who like chasing balls & learning treat rewards for rolling it. 5 Stars FUN & for puppies learning to problem solve. Fun will depend on dog's size, especially if they're mouthy (chew-wise) & how big they open--can the dog get the ball in its mouth to break it? Overall our experience is FUN=5 Stars
2) SAFETY: Our 5 month, 22" high, 30# Australian Pup spent 2 hours of continuous play, first time w IQ Ball. He could not hold or carry it in his small puppy mouth...this is not a problem but a good thing. Larger dogs wont necessary get the pt. if all they do is carry it around. Also---Reckon, any larger dog e.g. Germ. Shep., Rottie., Malamute & esp. Staffordshire Bull, Pit Bull Terriers, or larger mouth dogs could easily crush this lightweight plastic into shattered sharp bits. If the dog is easy going on toys it may be okay. Otherwise I'd worry about it breaking in the dog's mouth.
3) Cost to Time ratio is likely very good if, dog is not too "mouthy" with their toys. Rate it at a 3 overall for Med. to Large Dogs & 5 for pups & smaller dogs. Cost not worth it if dog crushes it in a minute.
Our little guy just started playing with IQ Ball again 5 hrs. after unpacking it. At first he chased it around the house for almost 2 hrs.
So Cost/Time used ratio depends on your dog's food motivation & love for balls that toss kibble out at various intervals. Set-up's a breeze. Delivery opening is adjustable, depending on kibble size & how much you want to reward the dog. We set delivery hole wide open the first 15 min. Then adjusted it smaller as he figured out, the more I roll this ball, the more treats come out.
SAFETY CAN BE AN ISSUE BUT OVERALL, WITH CERTAIN DOGS & DOG SIZE, THIS IQ BALL IS A GOOD 4 STARS. 1 other caveat: uneaten kibble left on floor can be a calling card for little "Mickies" who just LOVE dog kibble. We vacuumed a few times already.
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Reviewed in the United States on November 28, 2015
★★★★★ 5
Great for keeping dog busy and for rescues
Size: 3 INCH, Pattern Name: IQ TREAT BALL 3"
We got our dog from a shelter -- we don't know much about her history, but we imagine that she has never been socialized to play with toys because she has no interest in objects whatsoever. We've tried squeaky toys, chew toys, and ropes but to no avail. Other than her Kong (and she only plays with that because she wants the food; she has no interest in the Kong itself), this IQ Treat Ball is the only other toy that she plays with.
It took us a day and a couple of meals to teach her how to use it. In fact, we started out by making the toy a bit easier by removing the disk in the center altogether. The disk is still out, but we'll be sure to add it when we feel that she needs the challenge.
I LOVE the concept of this toy because it teaches dogs how to play, using food as a motivator, and allows them to really think and problem solve (especially for older rescue dogs who have never really been taught). My dog started out constantly getting the ball stuck under the couch and between furniture, but she now knows how to navigate around the house and how to avoid these "problem areas." She originally had no idea how to push the ball around, but now she noses it around eagerly for her kibble.
Aside from that, it is easy to open and easy to clean.
I see some negative reviews here, which is unfortunate because even though the toy is made of plastic, it IS well-made. (I should note here that my fiance and I have both dropped the ball from kitchen-counter height to the tile on multiple occasions because we're clumsy, and there are no dents or cracks from the impact.) Even on the packaging, the manufacturer gives you the warning that the ball is NOT for aggressive chewers and should absolutely only be given to a dog under guardian supervision. Giving the IQ Treat Ball to an aggressive chewer, not supervising him, and then blaming the manufacturer for injuries is irresponsible.
My dog is not a chewer, and I always watch her while she is playing with this toy, so I think that it's a great product. If you have a rescue/shelter dog who doesn't know how to play with toys, I think this is a fantastic start! I highly recommend!
UPDATE (7/15/2014): This toy has lasted a whole month since we started using it mid-June. We use it twice a day for breakfast and dinner, and it's still in good shape. The ridges around the ball do have some minor teeth marks on on them, but that's something that I expected would happen. As far as I'm concerned, it's still in great working order.
I am ecstatic with this toy because it has functioned as a segue for our rescue. As I mentioned above, she showed absolutely ZERO interest in toys before. However, we've slowly started getting her out of her anti-toy-socialization shell, and I think this IQ Treat Ball has served a part in that. We've had her for about two months, and in that time we've taught her out to play tug-of-war, how to chase after a ball (Very short distances for now, but hey! Baby steps, right?), and how to chew on ropes and other toys. Now, a lot of hard work has gone into shaping these behaviors on our part, but I really do think that the IQ Treat Ball has improved her appetite, not only for food, but for games as well.
My original verdict still stands -- I highly recommend!
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Reviewed in the United States on June 30, 2014
★★★★★ 5
Fun for your dog and fun for you to watch their antics with this Interactive IQ Treat Ball
Size: 3 INCH, Pattern Name: IQ TREAT BALL 3"
This delightful ball is both a feeding utensil and a workout all in one! Your dog not only eats its calories, it burns them off at the same time!
I have a little dachshund, Sophi, who loves her treat ball. They call it a treat ball but I use it for kibble that fits inside. Sophi doesn't need more treats than just a few every few days. If you fill this with treats, it's too much in my opinion, unless you have a really playful dog who needs to gain weight.
You place the treats or kibbles inside one half of the ball (the one without the hole in it). Then you adjust the size of the hole in the white center piece to allow either one or a few pieces to fall into the chamber at a time. You place the white flat part over top of the kibbles and then screw on the other half. There is a hole in the end of the ball and once the kibble/treats roll around in the chamber and make their way through the one hole inside to the opposite chamber and then reach that hole, they fall out. Sounds complicated but it's not.
This ball teaches the dog that rolling the ball makes treats/kibble fall out and they get to enjoy them. This is where IQ in the name comes in. It doesn't take long for a dog to figure out that they get treats by rolling the ball around and making them fall out.
Not only does the dog get exercise pushing the ball around your floor, it really is entertaining to watch. Our Sophi hears the kibbles fall out and if she doesn't immediately see them, because the ball continues to roll, we get to see that "where are they?" and the floppy dachshund ears frantically searching. We have hardwood floors so the ball rolling quickly and bumping into table legs and furniture, Sophi trying to get the kibbles--it's quite a show that has us laughing a lot.
It's a sturdy plastic ball, easy to fill and easy to put together. Small enough to tuck into a bag for travel--about the size of a baseball, or smaller. Will roll easy on carpet with a dog pushing it with their nose or very quickly on hardwood.
Rather noisy on hardwood floors with the assertiveness of the dog and it bumping into things, so watching something on TV can be difficult. But the entertainment you get, who needs TV?
It'll only hold about 1/2 c or a little more of small kibbles (you cannot pack it full) so it's more for multiple feedings, snacking or play. I ordered 2 of these so that when one was used and needed cleaned, there was always a clean one. I throw them in dish water or the dishwasher, top rack with no problem. No cracks or breaks in almost a yr of use.
Highly recommend this interactive treat ball. Not only will your dog enjoy it, you will enjoy watching your dog enjoy it.
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Reviewed in the United States on November 22, 2015
★★★★★ 5
Great Energy Outlet for an Active Dog!
Size: 4 INCH, Pattern Name: IQ TREAT BALL 4"
I have a 1 year old Australian Shepherd/Border Collie/Cattle Dog mix named Josie. When you look up "hyper-active dog" in the dictionary, you'll find her picture. She could run for days if she wanted to. I am always looking for things to help keep her from getting bored, and this treat ball is great!
She was only about 3 or 4 months old when I got this and introduced it to her. With a little help from me, she figured it out in no time. Now she's a pro, and knows exactly how to paw at the ball to make it spin and fling food out.
I have the 5-inch ball. Josie's not a big dog (40lbs) and she can get her mouth around the ball to pick it up. She only does this when it rolls onto the linoleum floor in the kitchen. I think it gets too loud for her (and me too!). I always put it away when the food is gone so she doesn't use it as a chew toy. She has never gotten bored of pushing it around and decided to chew on it. She can hear the food rattling around inside and continues to play with it until it is empty. This isn't the type of toy that you would give to your dog, and then leave them unsupervised.
The ball is made out of hard plastic, and does make a loud banging when it runs into walls, chairs, etc., but it has never left any marks on my walls or furniture. I actually prefer she use it on the carpet because it's quieter and she always gets all of the food pieces (it works just as well on carpet as it does on a hard floor, fyi). If I lived on the bottom floor or in my own house, I wouldn't have a problem with her using it on hard floors, however she always brings it back to the carpet on her own when it rolls off.
I like that the hole on the middle divider is adjustable, and you can also just take the middle part out of the ball so you can fit more food inside, the dog can see the food so they're more motivated to get it, or to make it easier for a younger/older dog to get food out.
My dog loves this ball. She gets excited when I take it out and start filling it with food. With the divider it has enough room for half of her portion. She always goes for the ball first, empties it, then to her dish. When she's done eating she always lays down and takes a nice long nap. I know that she enjoys it, and it definitely gives her 15-20 minutes of good, mentally-stimulating indoor exercise. It also taps into their prey-drive, and I think that helps make your dog feel like a dog.
Overall this is another great buy from Amazon. I did see it at the pet store (after I had gotten mine) for double the price!
***If you have a dog that is food-motivated and needs something to do, buy this. It's worth it.***
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Reviewed in the United States on October 26, 2011
★★★★★ 3
Great toy while it lasts
Size: 3 INCH, Pattern Name: IQ TREAT BALL 3"
We purchased one of these for our 10 pound Malshipoo as she gets bored easily and this thing definitely keeps her busy for awhile while she tries to get the treats out. She rolls and kicks it hard enough that I'm sometimes fearful it will break but after having owned it for 14 months it's actually held together quite nicely to my surprise. I think it came apart once and that's probably because I didn't have it screwed on correctly. The only reason I didn't rate it higher was two things: First, I don't like that the top section is in two parts and you have to fit them together just right for it to screw on. Why not make it one solid piece on the top? The second reason I didn't rate it higher is the white plate with the hole you twist to make the hole bigger or smaller is now a lot looser so when our dog bangs this against a wall or anything hard the hole shifts and makes it bigger thus the game doesn't last as long. So I'm purchasing a new one and hoping this doesn't happen again. I guess it's just wear and tear but I would have like it to last longer than 14 months. Maybe I guess my expectations aren't realistic. Hopefully the next one lasts longer.
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Reviewed in the United States on February 18, 2025
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