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Description
Rat gSAP 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 capture antibody against Gamma-Secretase Activating Protein (gSAP). 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 Gamma-Secretase Activating Protein (gSAP) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Gamma-Secretase Activating Protein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | γ-Secretase activating protein (gSAP), also known as protein pigeon homolog (PION), is a protein encoded by the gSAP gene. It regulates γ-secretase activity, specifically activating amyloid protein (APP) interaction to increase β-amyloid production without affecting the cleavage of other γ-secretase targets. The γ-secretase complex is an endoprotease that catalyzes the intracellular cleavage of integral membrane proteins. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates and other biological fluids |
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4.8 ★★★★★
Based on 10 reviews
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Product Reviews
★★★★★ 5
Worked for pantry
Size: 3Pack of 5 Tier (Not included planks), Size: 3Pack of 5 Tier (Not included planks)
These turned out perfect t for a pantry. Just fyi get better dry wall anchors and you’re golden! Definitely takes two people to put up tho!
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Reviewed in the United States on February 14, 2026
★★★★★ 5
Buy it.
This is not merely another guide to intensive care. Well-organized and detailed, it hits the right note between the things a beginner has to know (and probably has some idea about) and the things a beginner needs to know (but is clueless). It even includes a chapter on burnout. Recommended for everyone new to the ICU, and also everyone who has been around awhile. I’m going to get a lot of use from this text, I can already tell.
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Reviewed in the United States on June 19, 2018
★★★★★ 3
A little too beginner; doesn't translate well to USA patterns of practice
Format: Paperback
The book title really says it all, it really is the BEGINNER'S guide to the ICU for junior doctors and allied health professionals - more like an introduction to important concepts rather than a guide really. The strengths of the text come from its stated purpose of being a absolute, beginner's guide to critical care. The book would be appropriate for perhaps a 4th year med student or a intern who is very early in residency w/ little ICU experience or a newly minted APP; there's little to be gained by a advanced resident, fellow or practicing physician. The chapters are very short which provide a mere grazing-the-surface of important critical care concepts - some chapters are too short to really be useful (e.g. the paltry coverage of ultrasound in crit care (p. 159) is only 10 pages including pictures). The book, editors and authors are UK-based which makes the units of measurement, choice of drugs and some practice patterns, not consistent with what is typical in the USA.
For this reason I cannot recommend this text for American learners; e.g. blood glucoses are measured in mmol/L internationally, however USA, Germany use mg/dL where a normal BG in UK may be "4.4" but in the US one might consider a normal BG "80". This carries over again with concepts of ABG's and their utility in ventilator settings, respiratory emergencies and sepsis, etc. which become more confounding when using the PaCO2/PaO2 kPa instead of the mmHg used in American ICU's. When a BEGINNER is trying to learn the FUNDAMENTALS of crit care I recommend that a learner be introduced to the concepts using data measurement they are expected to utilize in practice rather than going through the mental gymnastics of doing conversions and THEN making a treatment decision.
The theme of UK and USA differences continues into drug therapy. For example when covering RSI and sedation the authors discuss the utility of sodium thiopental, however this drug has not been available in the USA for many years. In addition there were some other areas where some recommended drugs did not correlate w/ typical USA patterns and others that received hardly any mention (e.g. little mention of vasopressin as an adjunct in pressor support, other paralytics in RSI such as succinyl choline, rocuronium, CCB's and BB's in atrial fibrillation). Least of all there are multiple areas where drug/device names that refer to the same agent but would confuse a beginner starting in the USA (e.g. albuterol = salbutamol, aceteminophen = paracetamol, norepinephrine = noradrenaline, Guedel = OPA etc.).
Lastly, on the topic of UK vs worldwide differences the epidemiologic data mentioned refers to UK populations making it somewhat of an abstraction of the prevalence of disease in your area of practice if you're outside the UK. Which is fine, just be aware of that.
The chapters, however, are well organized and majority begin with a clinical case which I find is a approach that cements concepts in learner. If anything I feel that some are much to short, even for a beginner. I'm specifically referring to the Cardiac Arrythmias chapter (p 233). There is much to cover on this topic and the 5 pages dedicated to it is simply not enough and there is no further recommended reading. And importantly, the EKG figures were switched around on p234 and p235, which again does a beginning learner a disservice. I did find the chapters dedicated specifically to ICU concepts useful such as "Fighting the Ventilator" and "Endotracheal tube and tracheostomy problems" which cover just enough ground for the trainee. Unfortunately, none of the chapters have in-text citations with little primary references - I did have some questions regarding some chapter authors recommendations and I'm unable to look up where the works cited to review the quality of evidence.
There are multiple chapter authors and unfortunately this creates some redundancies. I could only find one area where there was a contradiction between authors which one author stated there is no contraindication for insertion of a NPA in setting of base-of-skull fracture (p.79) and on the next chapter another author stating that "nasopharyngeal airway is contraindicated if there is the possibility of a base of skull injury!" (p.87) - less than 10 pages apart. Again, there's no primary texts referenced and I can't confirm where the best, up to date evidence lies.
In SHORT: this is a useful text to the BEGINNER who is looking to obtain a broad overview of critical care CONCEPTS. It is pretty easy to read through and simple to digest where I a motivated learner could get through the full 440 pages relatively quickly and gain a good grasp & appreciation of the concepts of critical care. The text accomplishes its goal of being a BEGINNER'S GUIDE to ICU and explicitly identifies its target audience in the title: . . . . A Handbook for Junior Doctors and Allied Professional. I do NOT recommend the text to American trainees for the reasons above (drugs, units, differences in practice patterns) and I don't recommend the text to practicioners who have more experience.
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Reviewed in the United States on January 19, 2021
★★★★★ 3
Material
Format: Paperback
The material is not the greatest very basic and it is all UK based
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Reviewed in the United States on February 2, 2020
★★★★★ 5
Good
Format: Spiral-bound
Good quality book
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Reviewed in the United States on May 8, 2026