SKU: 5839112959

Mouse Rac1 ELISA Kit

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Description

Mouse Rac1 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.

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 Ras-Related C3 Botulinum Toxin Substrate 1 (Rac1) capture antibody. 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 Ras-Related C3 Botulinum Toxin Substrate 1 (Rac1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration.
Source Mouse
Synonym Mouse Ras Related C3 Botulinum Toxin Substrate 1 ELISA Kit
Detection Type Double antibody sandwich method
Composition
Name 9 6 T  match   set remark
Pre-coating 96 Well plate 8 Hole ×12 Strip without
Standard 2 branch
Dilute as per instructions
Universal diluent
2×20mL
without
Concentrated biotinylated antibody ( 100× )  
120uL
Dilute as per instructions
Concentrated enzyme conjugate ( 100× )
120uL
Dilute as per instructions
20× Washing liquid
2×10mL
Dilute as per instructions
Bottom thing ( TMB )
10mL
without
Stop liquid
6mL
without
Sealing film
4 Zhang
without
Instructions
1 Share
without
Background Ras-related C3 botulinum toxin substrate 1, also known as Rac1, is a protein encoded by the RAC1 gene. This gene can produce multiple alternatively spliced forms of the Rac1 protein, which appear to have distinct functions. It is a small signaling G protein (more specifically, a GTPase) and a member of the Rac subfamily of the Rho family of GTPases. Members of this superfamily appear to regulate a variety of cellular events, including control of GLUT4 translocation to glucose uptake, cell growth, cytoskeletal reorganization, antimicrobial cytotoxicity, and protein kinase activation. It is a pleiotropic regulator of many cellular processes, including the cell cycle, cell adhesion, motility, and epithelial differentiation. It is ubiquitously expressed in tissues and drives cell motility by forming lamellae. It is abundantly expressed in insulin-sensitive tissues, such as adipose tissue and skeletal muscle. It is also required for glucose uptake in skeletal muscle, which is activated by exercise.
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.
Storage Temp. If the unopened kit is stored at 4°C, the shelf life is 6 months.
Test Range 0.156-10 ng/mL
Applications Tissue homogenates, cell lysates, and other biological fluids
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Exchange/Return Notes
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SKU: 5839112959

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4.8 ★★★★★
Based on 7 reviews
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Product Reviews
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robert ferrell
Cuba, US
★★★★★ 5
Good
Color: Black, Size: 6 Panel
Love it , just what so needed , easy to put together (some what ) …
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 16, 2026
C
Verified Purchase
Crimm
Grantham, US
★★★★★ 3
Decent for the price. Instructions suck but most people should be able to figure it out. More below.
Color: Black, Size: 6 Panel, Color: Black, Size: 6 Panel
SUMMARY: 3 stars from me because it's firmly average. It's fine for the price. Assembly and materials are alright but I can see some caveats depending on your circumstances. Assembly instructions do a subpar job of pointing out some details. ASSEMBLY INSTRUCTIONS: Maybe it's because I'm autistic and/or building model kits and assembling stuff is my jam, but I honestly thought people were exaggerating when they were complaining about the instructions and I'd be able to flex my ~superior assembly skills.~ I was wrong! These instructions genuinely suck, and whoever is responsible for making them should be ashamed. The instructions do a poor job of calling out some details regarding orientation of parts, and some of the images do not actually match the physical parts. For example, it does not really outline the assembly of the end panels clearly, and I can see someone accidentally using the wrong poles. I've drawn over a photo to show what you should do to try and make it clearer. Additionally, the manual shows a flathead screw for bolting the feet into position, but the actual screw is not a flathead. It also does not point out the counterbore, so if you aren't paying attention you may put the foot pads onto the wrong side of the foot. It's also missing the fact that you need to use another one of the plastic pieces when you finish assembling a panel. ACTUAL ASSEMBLY & MATERIALS: To their credit, all of my bags were clearly labeled. The assembly process wasn't difficult. It's mostly just tedious and requires a fair amount of space. I was able to assemble it by myself without any real difficulty. However, the way it's assembled means two things. One, the fabric parts aren't removable without disassembly. So if you want to use this in an environment where they would require cleaning, I would seriously recommend looking for another option. Or, you could buy this just to use the frame pieces and then somehow buy or make your own fabric pieces designed to be removable with velcro or something. Two, because of the materials I really don't have a lot of faith in this thing surviving disassembly and reassembly. Like a lot of sorta-cheap-but-convenient furniture, it uses those spring-button connections and plastic inserts with self-tapping screws. Those things are not really meant to be disassembled and rethreaded. It also relies a lot on the tension of the poles and the fabric to keep everything rigid and squared, which I think puts a lot of pressure on the aforementioned buttons, plastic inserts, and the hollow metal rods. So I feel like that will also cause issues with disassembly and reassembly. Basically once this thing is assembled, it's not really meant to be disassembled. The best you can do is spot-clean the fabric if you need to. Speaking of the fabric, I didn't see any labels on them or anything in the manual that says what they are, but they feel like some kind of polyester. They generate static electricity pretty easily, and pet hair and debris sticks easily. So that's another downside of them not being easily removable. For the most part it does seem pretty stable. The poles seem to be pretty uniform in length so they're all making contact with my floor. Obviously this isn't structural so it shouldn't be supporting anything, but the two main feet seem to be doing fine with keeping this thing upright. CLOSING THOUGHTS: Really, it's fine for what it is, but it could be better in a lot of little ways and the substandard quality of the instructions just seems unprofessional to me, which is why I'm being so harsh with my rating. Depending on your needs and environment you may want to consider a different option. Preferably one that is made to be disassembled with better materials, and/or one with fabric pieces made to be removed easily.
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Reviewed in the United States on May 28, 2023
B
Verified Purchase
Barb
Dallas, US
★★★★★ 1
Not as it seems
Color: Black, Size: 6 Panel
these are awful... Each individual panel is fine, but when you put it together it can barely stand up, and the clips that hold it together keep popping off. Steer clear of this item.
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Reviewed in the United States on April 7, 2026
T
Verified Purchase
Timothy Beckner
Phoenix, US
★★★★★ 5
As described
Color: White, Size: 6 Panel
Works great in my clinic
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 17, 2026
D
Verified Purchase
Debbie Flynn
Massapequa, US
★★★★★ 5
Love it it was perfect for what I needed it for. Good quality!
Size: 4 Panel, Color: Black
It like a very dark chocolate brown/black. What I liked most about it is that you just pull it out of the box take the long plastic film off of it and it's ready to go. It has protective feet on the bottom. It's very attractive and it was so easy. It's so nice when you have a product you don't have to assemble.
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Reviewed in the United States on May 13, 2026

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