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Description
Mouse GRa 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 Glucocorticoid Receptor Alpha (GRa) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Glucocorticoid Receptor Alpha (GRa) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Glucocorticoid Receptor Alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The glucocorticoid receptor (GR) is a member of the conserved nuclear receptor superfamily and a nuclear transcription factor. GRs include two subtypes: the normal GRα and the variant GRβ. GRα is the result of different splicing of the same transcript of the GR gene. At the mRNA level, GRα comprises exons 1 to 8, including exon 9α. It is expressed in nearly all tissues and cells, and glucocorticoids primarily exert their biological effects by binding to GRα. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.1 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 5
All Five Toys Are Mine Now — A Review by Patches the Pomeranian
Color: Pink, Color: Pink
⭐ “Hi, I’m Patches, and These Toys Are All Mine Now” — A Review by a Very Busy Pomeranian ⭐
Hello, my name is Patches, and I am what the humans call a “toy enthusiast.”
What that really means is: if a toy enters this house, it belongs to me immediately, no questions asked.
The Pack Leader brought home this 5‑Pack Puppy Chew Toy Set — a pea pod, a wing‑shaped chew, and three cute fabric‑tied toys. Five toys. Five opportunities for greatness. Five items for my personal collection.
He tried to hand one toy to each of us like we’re participating in some kind of polite sharing ceremony.
Very sweet. Very unrealistic.
The moment my sisters glanced away for a single second, I activated my elite move:
The Pomeranian Slide‑In‑and‑Snatch.
I took my toy.
I took their toys.
I took the toys on the floor.
I would’ve taken the mail if it looked interesting.
Now I have all five toys, and my sisters have… well… the joy of watching me parade around with them like a tiny fluffy champion.
🦴 My Professional Toy Review
Pea pod: Chews beautifully and looks adorable when I carry it around like a prize.
Wing bone: Excellent for dramatic chewing sessions.
Fabric toys: Perfect for shaking, tossing, and showing off.
They don’t squeak, they don’t bounce — and honestly, that’s perfect.
I enjoy a quiet chew while plotting my next toy acquisition.
😬 Teething & Dental Stuff
The humans say these toys help clean teeth and massage gums.
I don’t know about all that.
I just know they feel good to chew and keep me busy while my sisters stare at me in disbelief.
🎁 Final Thoughts
If your puppy enjoys chewing, collecting, or claiming ownership of everything within paw‑reach, this set is a fantastic choice.
5/5 stars.
Would absolutely claim again.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 28, 2026
★★★★★ 5
Anya loves her new baby toys
Color: Pink, Color: Pink
This review is for the YZHZIIR 5 Pack Puppy Chew Soft Rubber Dog Toys.
As you can see in my photos, my Borzoi “puppy” is quite large. I knew these toys are not meant for extra large breed puppies, but for some odd reason, ours loves little tiny toys. She usually will just carry them around in her mouth and will gently play with them. That being said, she will have these baby toys in her “supervised only” drawer.
This order came with 5 toys per pack, as stated on line. They look exactly like what you see, and it even specifies the sizes. I think if you have a toy breed, these are suitable since they’re safe to chew (not ingest). They’re BPA and phthalates free, made of natural rubber, so I’m not concerned about her developing a third eye or a lump somewhere on her body.
My puppies are the apple of my eye, so they are supervised whilst playing with small, easy to tear apart toys or goodies that may have a choking hazard. So my recommendation is that unless you are keeping a close eye on them, stick to giving these to toy breeds with tiny teeth, or very young puppies who could barely chomp down on soft puppy food.
The price is unbeatable. For 5 toys, you pay only $9.99!
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Reviewed in the United States on June 12, 2026
★★★★★ 1
Made Puppy Sick!
Color: Blue, Color: Blue
Fell apart fast, my pup tore pieces off and it made him sick. I really wanted these to work because they are so cute as "baby" toys for puppies and the listing promises “durable & chew‑resistant” toys that can stand up to sharp puppy teeth and even says they’re safe for extended chewing with no health risks because they’re made from “100% natural rubber” with no BPA or harmful chemicals. Unfortunately, that wasn’t my experience at all. My pup had these for only a short time before he chewed pieces off, and the toys started breaking apart way faster than expected.
After swallowing some of the pieces, he ended up sick to his stomach, which meant the toys had to go straight in the trash. He was up all night with diarrhea and vomiting, at first, I didn't put it together until I found blue pieces of the rubber toy in his vomit.
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Reviewed in the United States on June 5, 2026
★★★★★ 3
good toys but better for gentle / non aggressive teethers
Color: Blue
cute but not very durable. better for dogs who are not aggressive chewers even though they are young pups.
when they are broken apart by teeth, the pieces are at least clean cuts. no big concerns of ingestion
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 7, 2026
★★★★★ 5
Cute set for my 2-1/2 pound chihuahua puppy, and this set would also make a great gift
Color: Pink, Color: Pink
First off, I like how each piece is individually wrapped and then placed all together in a plastic bag. While it's not in a gift box, I feel like this would still make a really cute gift (that shows up in good shape, too.)
For the current price of under ten bucks, this feels like a really great value for five cute toys.
My chihuahua puppy is only 2-1/2 pounds, so some of these are a bit larger for her, but that's to be expected at her tiny size, and it doesn't stop her from enjoying them.
These feel really sturdy to me for smaller dogs to chew on. I personally love the added "ribbons" because I think they're adorable and because my dog loves things like that. They really get her to engage with toys even more. while also giving me another way to interact with them more as I play with her.
I could see how those fabric pieces may wear down over time for a larger dog, but I think it depends on how much a dog focuses on chewing on those pieces. Fortunately, the plastic toys will still be cute without those fabric pieces if anything happens to them.
Unfortunately, the plastic pieces have a very slight odor. I'm super sensitive to smells, so you may not notice it at all. It's certainly not as bad as so many other plastic items, and it came right off when I washed it in dish soap (which I would do before giving these to my pup anyways.)
I hope your small dog enjoys these as much as we do, and I hope you find this review helpful!
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Reviewed in the United States on May 27, 2026
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