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Mouse IFN-γ ELISA KitProduct Specification Usage Need to bring your own test equipment 1. Microplate reader (can measure the absorption value of 450nm detection wavelength and 540nm or 570nm correction wavelength) 2. High precision liquid dispenser and disposable suction head 3. Distilled water or deionized water 4. Washing bottle (spray bottle), multi channel plate washer or automatic plate washer 5. 500mL cylinder One, preparation before the experiment 1. Sample
Product Specification
| Usage |
Need to bring your own test equipment 1. Microplate reader (can measure the absorption value of 450nm detection wavelength and 540nm or 570nm correction wavelength) 2. High precision liquid dispenser and disposable suction head 3. Distilled water or deionized water 4. Washing bottle (spray bottle), multi-channel plate washer or automatic plate washer 5. 500mL cylinder One, preparation before the experiment 1. Sample collection and storage ① Cell culture supernatant: particles should be removed by centrifugation; Test the samples immediately. If the sample is not tested in time after collection, it is recommended that the sample be divided according to the dosage and stored in the refrigerator at -20 ° C to avoid repeated freezing and thawing. Samples may need to be diluted (1×) Dilute. ② Serum: Samples were collected using a serum separation tube (SST) and samples were left at room temperature for 30 minutes. The samples were centrifuged at 1000g for 15 min. Serum was immediately removed and tested immediately. If the sample is not tested in time after collection, it is recommended to repack according to a single dosage and freeze in ≤ -20℃ refrigerator to avoid repeated freezing and thawing. Samples may need to be diluted (1×) Dilute. ③ Plasma: Plasma was collected using EDTA, heparin, or citric acid as an anticoagulant, centrifuged at 1000g for 15 min within 30 min of collection, and tested immediately. If the samples are not detected in time after collection, it is recommended to separate the samples according to the single dosage and freeze them in &le. -20℃ refrigerator to avoid repeated freezing and thawing. Samples may need to be diluted (1×) Dilute. 2 Reagent preparation (Place all reagents and samples at room temperature for 15 minutes before use. It is recommended that all experimental samples and standards do double hole detection ) 1× Preparation of washing solution: concentrated washing solution in the kit is 20× Mother liquor, diluted to 1× with distilled water before use; Working liquid. Example: Take 10mL concentrated washing solution +190mL distilled water to 200mL, the actual operation can be calculated first, then make up. ②1× Dilution with buffer preparation: concentrate dilution in the kit with buffer 10× Mother liquor, diluted to 1× with distilled water before use; Working solution. example: Take 3mL of concentrated dilution with buffer +27mL of distilled water to a constant volume of 30mL. In practical operation, the required dilution buffer can be calculated according to the dilution multiple of the sample, and then the preparation can be made. ③ Detection of antibody: the dry powder was centrifuged to the bottom of the tube, and 110uL dilution buffer (1×) was used. Dissolve and let stand at room temperature for 5 minutes to obtain 100× Mother liquor; Dilute to 1× before use; Working solution. Calculate the desired volume by using 100uL per well. example: 10 Wells were used, then take 10uL of 100 times the working concentration of the test antibody, using dilution buffer (1×) Constant volume to 1mL, get 1mL of 1× The working concentration of the detected antibody. ④SA-HRP: SA-HRP is 40× Mother liquor, use dilution buffer before use (1×) Dilute to make 1× Working solution, 100uL required per well. example: used 10 holes, then take 25uL of 40× Mother liquor +975uL dilution buffer (1×) Constant volume to 1mL to obtain 1× of 1mL; The working concentration of the detected antibody. ⑤ Chromogenic agent: according to 100uL per well, calculate the amount needed for this test, take out the corresponding volume of chromogenic agent, avoid light; The removed chromogenic agent is only used on the same day. ⑥ Standards: lyophilized standards with dilution buffer (1×) Redissolve, redissolve volume 1000uL, to obtain a concentration of 2000pg/mL standard mother liquor. Gently shake for at least 5 minutes, it is fully dissolved. Add 300uL dilution buffer (1×) to each dilution tube. . The standard mother liquor is diluted according to the picture below, and each tube must be fully mixed before pipetting to the next tube. The standard mother liquor without dilution can be used as the highest point of the standard curve (2000pg/mL), and the dilution buffer (1×) Can be used as a standard curve zero (0 pg/mL). 2, operation steps 1. Prepare all required reagents and standards; 2 Remove the microplate from the sealed bag that has been balanced to room temperature, and put the unused slat back into the aluminum foil bag and re-seal it; 3. Add 300uL washing solution to the microplate, let it soak for 30 seconds, discard the washing solution and pat the microplate dry on absorbent paper, please use immediately do not let the microplate dry; 4. Add different concentrations of standard, experimental samples or quality control into the corresponding Wells, 100uL for each well. The Wells were sealed with plate adhesive and incubated at room temperature for 2 hours. 5 Suck the liquid out of the plate and wash the plate using a bottle washer, a multi-channel plate washer, or an automatic plate washer. Add 300uL of washing liquid to each well, and then suck the washing liquid out of the plate. Repeat 3 times. Every time you wash the plate, try to absorb the residual liquid to help you get a good test result. At the end of the last plate wash, please blot all the liquid in the plate or invert the plate and pat all the residual liquid in the absorbent paper; 6. Add 100uL detection antibody to each microwell. Seal the reaction Wells with sealer tape and incubate for 2 hours at room temperature; 7. Repeat the plate washing operation of step 5; 8. Add 100 ULSA-HRP to each microwell and incubate for 20 minutes at room temperature. Be careful to avoid light; 9. Repeat step 5 to wash the plate; 10. Add 100uL of color development solution to each microwell, incubate at room temperature for 5-30 minutes, pay attention to avoid light; 11. Add 50uL of termination solution to each microwell, and the color of the solution in the well will change from blue to yellow. If the color of the solution turns green or the color change is inconsistent, tap the microplate to mix the solution evenly; 12. Within 30 minutes after the termination solution is added, the absorbance value at 450nm is measured using a microplate reader and 540nm or 570nm is set as the correction wavelength. If the dual wavelength correction is not used, the accuracy of the results may be affected; 13 Calculation results: The corrected absorbance values (OD450-OD540/OD570), the compound reading were averaged for each standard and sample, and then the average zero standard OD value was subtracted. Standard curves were created by 4-parameter logic (4-PL) curve fitting using computer software. Alternatively, a curve can be generated by plotting the logarithm of the concentration of the standard against the logarithm of the corresponding OD value, and the best fit line can be determined by regression analysis. This process produces an adequate but less accurate fit to the data. If the sample is diluted, the concentration should be multiplied by the dilution.
Note: The standard curve data provided is for reference only. The sample content should be calculated according to the standard curve drawn in the same test.
3. Kit parameters 1. Recovery: Different levels of mouse IFN-&gamma were incorporated into the cell culture medium samples; And the recovery rate was determined. The recovery ranges from 82 to 110%, with an average recovery of 94%. 2. Sensitivity: mouse IFN-γ The minimum detectable dose (MDD) of murine IFN - & gamma was generally less than 1.8pg/mL. The lowest detectable value is the corresponding concentration calculated from the mean of the zero absorbance values of 20 standard curves plus two standard deviations. 3. Calibration: The ELISA kit was prepared with highly purified recombinant mouse IFN-&gamma expressed in E. coli. Corrected by protein. 4. Linearity: four different samples were mixed with high concentrations of mouse IFN-γ , and then with diluent (1×) Measure linearity by dilating the sample to within the detection range. 5. Specificity: The ELISA could detect both native and recombinant mouse IFN-γ Protein. Combine the following factors with diluent (1×) Formulated to a concentration of 50ng/mL to detect the difference with mouse IFN-γ The cross reaction. 50ng/mL interfering factor was incorporated into the intermediate range of recombinant mouse IFN-γ To detect the effect of IFN-&gamma on mice; The interference of IFN - & gamma. No significant cross-reaction or interference was observed. 4, common problem resolution 1. The white board (no color), after the completion of color
2. Flower plate (blank, negative and positive controls were normal, but the OD value of sample Wells was significantly higher)
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| Theory | This kit with double antibody sandwich enzyme-linked immunosorbent detection technology. Specific anti-mouse IFN-γ antibody was precoated on a high-affinity microplate. Enzyme label plate hole with standard substance, the sample under test and biotinylated antibodies detection, after incubation, samples of IFN - gamma combined with solid phase antibody and detection antibody, immune complex. After washing to remove not combined with material, by adding horseradish peroxidase labeled chain mildew avidin (Streptavidin - HRP). After washing, adding chromogenic substrate, dark color. The reaction was terminated by adding the termination solution, and the absorbance value was measured at 450nm wavelength (reference correction wavelength 540nm or 570nm). | ||||||||||||||||||||||||||||||||||||||||||||||||
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| Background | Interferon-γ (IFN-γ, also known as type II interferon) is an important immunomodulatory cytokine, which has been found to have antiviral activity. IFN-γ plays a key role in the host defense process through its antiviral, antiproliferative and immunomodulatory functions. "In many cell types, IFN-γ induces cytokine production and upregulates the expression of a variety of membrane proteins, including types I and II major histocompatibility complex (MHC) antigens, Fc receptors, leukocyte adhesion molecules, and B7 family antigens." IFN-γ is a potent activator of macrophage response. It directs the synthesis, conversion, and secretion of immunoglobulin by B cells. IFN-γ affects the development of T helper cell phenotype by inhibiting the differentiation of Th2 cells and stimulating the development of Th1 cells. IFN-γ plays an important role in the progression of autoimmune diseases and inflammatory diseases such as atherosclerosis. bioactive IFN - gamma is made up of two non covalently linked with different degree of glycosylation of 20 to 25 kda homologous dimer composed of subunits. Mature mouse IFN-γ shared 86% amino acid sequence identity with rat IFN-γ, and 38-44% amino acid sequence identity with bovine, dog, cotton rat, horse, cat, human, pig and rhesus monkey IFN-γ. The IFN-γ dimers assemble before the transmembrane IFN-γRI(α subunit) and then bind to the transmembrane IFN-γRII(β subunit), thus forming an active receptor complex containing two α and β subunits. IFN-γRII in the receptor complex increases ligand affinity as well as the efficiency of signal transduction. Although the α chain is widely expressed in a variety of cell types, the receptor β chain is associated with the state of IFN-γ response and its expression is tightly regulated. Under inflammatory conditions, IFN-γ can be produced by a variety of cell types, including dendritic epidermal /γδT cells, keratinocytes, peripheral γδT cells, mast cells, neurons, CD8+T cells, macrophages, B cells, neutrophils, natural killer cells, CD4+T cells, and testicular spermatids. |
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| General Notes | 1. Please use the kit within the validity period. 2. Components of different kits and different batch kits should not be mixed. 3. If the sample value is greater than the highest value of the standard curve, the sample should be diluted with (1×) The samples were diluted and retested. "If the cell culture supernatant sample needs to be diluted in a distributed manner, cell culture medium may be used for intermediate dilutions, except for the last step when diluent is used." 4. The difference of the test results can be caused by a variety of factors, including the operation of the experimenter, the use of the pipettor, the washing technique, the reaction time or temperature, the storage of the kit, etc. 5. The termination solution in the kit is acidic. Please protect your glasses, hands, face and clothes when using it. 6. For scientific research only, not for in vitro diagnosis. |
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| Storage Temp. | Unopened kit, 2-8° C Storage | ||||||||||||||||||||||||||||||||||||||||||||||||
| Test Range | 31.3pg/mL-2000pg/mL |
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4.6 ★★★★★
Based on 22 reviews
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Product Reviews
★★★★★ 5
A must-read for professionals seeking to stay ahead in a rapidly evolving landscape.
Format: Kindle
“ChatGPT for Cybersecurity Cookbook” by Clint Bodungen offers a well-organized, practical deep dive into the integration of generative AI, specifically ChatGPT, within cybersecurity. From the perspective of a cybersecurity professional, this book stands out as a valuable resource for anyone looking to modernize and automate many of the repetitive tasks in cybersecurity, such as threat analysis, vulnerability management, and policy creation.
The book’s greatest strength lies in its focus on practical, actionable “recipes.” Each chapter walks you through real-world use cases for applying ChatGPT to cybersecurity operations. For instance, Bodungen effectively demonstrates how to automate penetration testing and vulnerability assessments using AI-driven techniques, something that can significantly streamline the red team’s workflow . The book’s use of LangChain for parsing and analyzing vulnerability reports, as well as integrating AI for governance, risk, and compliance (GRC) assessments, provides blue teams with a powerful set of tools for enhancing their day-to-day operations .
One of the highlights is how the author tackles threat intelligence, using ChatGPT alongside the MITRE ATT&CK framework, which can help automate complex tasks like identifying and categorizing threat actors and their tactics, techniques, and procedures (TTPs). For any seasoned cybersecurity analyst, this can save a lot of time and reduce human error when managing large volumes of data .
However, it’s important to note that the book assumes a decent understanding of Python and core cybersecurity concepts. It’s not for absolute beginners but provides significant value for professionals looking to integrate AI into their operations more efficiently.
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Reviewed in the United States on October 17, 2024
★★★★★ 5
A Career-Changing Read for Cybersecurity Professionals
Format: Paperback
This book completely changed the trajectory of my career. As a self-taught security professional with 30 years of experience, I found myself feeling stagnant and losing interest in cybersecurity. The rapid changes in the field made me question my next steps, and I had zero knowledge of Generative AI, let alone its applications in cybersecurity.
Then, the week this book was released, I bought it—and told no one. It was my personal experiment, a chance to dive into something new. Since then, I’ve read multiple books on Generative AI for Cyber, applied the concepts, and expanded my expertise.
Today, I am considered the SME for Cyber AI in my organization, actively participating in AI working groups, and regularly speaking, mentoring, and presenting on the topic.
In fact, I was the third reviewer of this book (5 stars) and deleted the review after about a week. I didn’t want someone who knew me to connect the review to me and figure out how I got so smart on the topic so quickly. Now, I’m back, giving it the review it truly deserves.
This book reignited my passion for cybersecurity and relaunched my career. If you're in cyber and looking for a way to break through stagnation or explore the future of AI in security, this is the book that could redefine your path—just like it did for me.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 4, 2025
★★★★★ 5
Clint is the Real Deal and this is an Excellent Book!
Format: Paperback, Format: Paperback
Clint knows his stuff, and I’ve also taken his accompanying training course. This book will give you some insights in how to apply a variety of techniques around ChatGPT and other engines and how to use them effectively in your job. Highly recommended!
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Reviewed in the United States on June 21, 2024
★★★★★ 3
Happy to have the book - damaged though
Format: Paperback, Format: Paperback
Excited to dive into this book but don’t love that it’s damaged.
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Reviewed in the United States on October 19, 2024
★★★★★ 5
Great GPT reference
Format: Paperback
Everyone I know is seeking the best options to build AI/ML, and GPT solutions into their current offerings. The “ChatGPT for Cybersecurity Cookbook” (Packt, 2024) by Clint Bodungen offers a detailed schematic for building effective ML solutions into existing practices. The book includes ten chapters, each focusing on a different area. The areas demonstrate clear solutions for framing a solution, building effective tools and library interactions, explaining what the solution accomplishes, and offering advanced solutions. Recommend the book for anyone working to improve their ability to use ChatGpt in accelerating cybersecurity solutions.
Each section approaches a different solution but the overall format is an outstanding deep dive for each section. The analysis starts with an introduction to the roles and formats. This details how to construct the ChatGPT query with enough guidance to answer the desired question. The next step suggests how the AI/ML tools find the right answers. The last section in every chapter details a “there’s more” showing how to expand the different questions. Chapters include building ChatGPT into Python language and API calls so one can not just launch a query, but have the query launch every time code executes.
All the chapters follow the same path but with different answers. This cookbook shows how roles and queries can be modified to deliver what one wants. One of the biggest benefits is seeing role construction from a cybersecurity expert with 20 years of governance experience to a threat analyst who desires to incorporate MITRE’s ATT&CK frameworks. The book demonstrates how to dig into the various sections for success. Some of the later sections demonstrate how one can point ChatGPT at specific data factors, for example, the AI tool can not execute commands to find traffic data but can be pointed to already collected traffic data to summarize answers.
One key flaw to remember is ChatGPT does not solve security problems for you. The best solutions point to frameworks and potential answers. The frameworks still need to be executed by a team or individual to create effective software models, it is just a step ahead in the overall process. That same framework really shows up in the threat analysis, training, and incident response sectors. Each solution builds a format to suggest some answers if pointed at the correct data. The answers are likely to be accurate but one still may need to verify against existing data and metrics. The other gap maybe if the ChatGPT tool is not pointed at specific information, answers may be skewed to what it can achieve.
Overall, if you want a quick way to incorporate an AI/ML tool, then “ChatGPT for Cybersecurity Cookbook” provides an excellent reference. From the cookbook perspective, one gets all the recipes for various security items, what the finished product should look like, and potential areas to improve in the future. The explanations are all geared to cybersecurity Recommend this as a key reference for anyone working with ChatGPT functions.
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Reviewed in the United States on June 11, 2024