Rat IL-1β/IL-1F2 ELISA Kit
Rat IL-1β/IL-1F2 ELISA Kit
Price:
Regular price
$450 USD
Regular price
Sale price
$450 USD
Unit price
per
For shipping services or bulk orders, you may request a quotation.
Secure checkout with
View full details
Product Details
Product Details
Product Specification
| Usage |
1. Microplate reader (measurable 450nm Absorption value of detection wavelength and 540nm Or 570nm Absorption value of corrected wavelength) 2. High precision liquid dispenser and disposable tip 3. Distilled or deionized water 4. Bottle washer (spray bottle), multi-channel plate washer or automatic plate washer 5. 500mL 1. Preparation before the experiment 1. Sample collection and storage ① Cell culture supernatant: particulate matter should be removed by centrifugation; Test the sample immediately. If the sample is not tested in time after collection, it is recommended to pack it according to the amount used once and store it frozen in -20℃ In the refrigerator, avoid repeated freezing and thawing. The sample may need to be used with a diluent ( 1× ② Serum: Use serum separation tubes ( SST ) Collect samples and place samples at room temperature 30 Minutes. Centrifugation 15 Minutes, with a rotation speed of 1000g ≤-20℃ In the refrigerator, avoid repeated freezing and thawing. The sample may need to be used with a diluent ( 1× ③ Plasma: Use EDTA , heparin or citric acid as an anticoagulant to collect plasma, after collection 30 Centrifuge within minutes 15 Minutes, with a rotation speed of 1000g ≤-20℃ In the refrigerator, avoid repeated freezing and thawing. The sample may need to be used with a diluent ( 1× ) dilution. 2. Reagent Preparation ( Please place all reagents and samples at room temperature before use and let them stand 15 Do repeat hole detection ） ①1× Preparation of washing solution: The concentrated washing solution in the kit is 20× Mother liquor should be diluted with distilled water before use to 1× Example: Take 10mL Concentrated wash +190mL Distilled water to volume to 200mL ②1× Preparation of buffer for dilution: The concentration and dilution buffer in the kit is 10× Mother liquor, dilute with distilled water before use to 1× Working fluid. Example: 3mL Buffer for concentration and dilution +27mL Distilled water to volume to 30mL 。 In actual operation, the required amount of dilution buffer solution can be calculated according to the sample dilution factor, and then prepared. ③ Antibody detection: Centrifuge the dry powder to the bottom of the tube and use 110uL 1× ) Dissolve and let stand at room temperature 5 Get after minutes 100× Mother liquor; Before use, dilute to 1× Working fluid. According to the amount per well 100uL Example: Used 10 Hole, then take 10uL Of 100 1× ) Constant volume to 1mL , obtained 1mL Of 1× Detection antibody at working concentration. ④SA-HRP SA-HRP For 40× Mother liquor, use dilution buffer before use ( 1× ) Diluted and formulated 1× Working fluid, the required amount per hole is 100uL Example: Used 10 Hole, then take 25uL Of 40× +975uL Buffer for dilution ( 1× ) Constant volume to 1mL , obtained 1mL Of 1× ⑤ Color development solution: per well 100uL ⑥ Standard: Dilution buffer for lyophilized standard ( 1× 1000uL , obtaining a concentration of 4000pg/mL Standard mother liquor. Gently shake at least 5 Minutes, it is fully dissolved. Add to each dilution tube 300uL Buffer for dilution ( 1× 4000pg/mL ), buffer for dilution ( 1× ) can be used as a standard curve zero ( 0pg/mL ）。  2. Operation steps 1. Prepare all necessary reagents and standards. 2. Take out the microplate from the sealed bag that has been equilibrated to room temperature. Please put the unused slats back into the aluminum foil bag and re-seal; 3. Add to the microplate 300uL Washing liquid, let stand and soak 30 Seconds, discard the lotion and pat the microplate dry on absorbent paper. Please use it immediately and do not let the microplate dry; 4. Add different concentration standards, experimental samples or quality control articles into the corresponding wells, and each well 100uL 。 Seal the reaction wells with plate sealing tape and incubate at room temperature 2 hour ; 5. Suck the liquid in the plate and wash the plate using a washing bottle, a multi-channel plate washer or an automatic plate washer. Washing solution per well 300uL Then the wash liquid in the plate is aspirated off. Repeat Operation 3 6. Within each well added 100uL 2 Hour; 7. Repeat th 5 Step washing operation; 8. Within each well added 100uLSA-HRP 20 Minutes. Be careful to avoid light; 9. Repeat th 5 Step washing operation; 10. Within each well added 100uL 5-30 Minutes, pay attention to avoiding light; 11. Within each well added 50uL Stop solution, the color of the solution in the well will change from blue to yellow. If the color of the solution changes to green or the color changes are inconsistent, pat the microplate gently to mix the solution evenly; 12. After addition of stop solution 30 Within minutes, measured using a plate reader 450nm Absorbance value, set 540nm Or 570nm As a correction wavelength. If dual-wavelength correction is not used, the accuracy of the results may be affected; 13. Calculation results: Add the corrected absorbance values of each standard and sample (OD450-OD540 OD570) , average of repeated well readings and then subtract the average zero standard OD Value. Using computer software for four-parameter logic (4-PL) Curve fitting creates a standard curve. Another way is to plot the standard concentration and make the logarithm with the corresponding OD The values were logarithmic to generate a curve, and the best fit line was determined by regression analysis. This process can generate a data fit that is sufficiently useful but less accurate. If the sample is diluted, the concentration should be calculated by multiplying the dilution factor.  3. Kit parameters 1. Recovery: Different levels were spiked into cell culture medium samples Rat IL-1β/IL-1F2 The recovery rate was determined. The recoveries range from 82-110% , the average recovery was in 94% 。 2. Sensitivity: Rat IL-1β/IL-1F2 The lowest measurable dose ( MDD ) is generally less than 1.8pg/mL 。 The lowest measurable value is determined according to 20 The corresponding concentration is calculated by adding two standard deviations to the mean value of the zero-point absorbance values of each standard curve. 3. Linearity: 4 High concentrations of different samples were incorporated into Rat IL-1β/IL-1F2 , followed by a diluent ( 1× ) Dilute the sample to the detection range and determine its linearity. 4. Specificity: This ELISA Method can detect natural and recombinant Rat IL-1β/IL-1F2 Egg whites. The following factors were mixed with diluent ( 1× ) configured to 50ng/mL Concentration to be detected with rats IL-1β Cross-reactivity of. Will 50ng/mL Interference factors incorporated into the intermediate range of recombinant rats IL-1β In the control article, to detect the effect on rats IL-1β Of interference. No significant cross-reactivity or interference was observed.
4. Analysis of frequently asked questions 1. Whiteboard (no color appears after color rendering is complete)
2. Flower plate (blank, negative positive control normal, but specimen well OD Values are significantly higher)
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Species Reactivity | Rat | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Theory | This kit adopts double antibody sandwich enzyme-linked immunosorbent detection technology. Specific anti-rat IL-1beta/IL-1F2 antibodies were pre-coated on high affinity enzyme plates. The standard substance, the sample to be tested and the biotinylated detection antibody are added to the well of the enzyme label plate, and after incubation, the IL-1beta/IL-1F2 present in the sample binds to the solid phase antibody and the detection antibody to form an immune complex. After washing to remove unbound material, horseradish peroxidase-labeled Streptavidin-HRP was added. After washing, a chromogenic substrate is added to protect the color from light. A stop solution was added to stop the reaction, and the absorbance value was measured at a wavelength of 450 nm (reference calibration wavelength of 540 nm or 570 nm). | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Synonym | catabolin, IL1 beta, IL-1 beta, IL-1, IL1B, IL-1b, IL1-beta, IL-1F2, IL1F2IL-1 beta, interleukin 1, beta, interleukin-1 beta, preinterleukin 1 beta, pro-interleukin-1 beta, Rat interleukin 1β Elisa kit | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Composition |
Please use within the expiration date of the kit
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Background | Interleukin -1 （ IL-1 ) Protein family members include classical IL-1 And IL-1 。 IL-1 Receptor antagonists ( IL-1RA ）、 IL-18 、 IL-33 、 IL-1F5-F10 、 IL-1 And IL-1 Have the same cell surface binding receptors and form the same biological function. Unstimulated cells in healthy people do not produce IL-1 The only exceptions are skin keratinocytes, some epithelial cells, and some cells of the central nervous system. But in response to inflammatory agents, infection, or microbial endotoxin, IL-1 Expression in macrophages and other types of cells increased dramatically. IL-1 In immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, GH/IGF-1 Plays an important role in the physiological process of. Multiple diseases and IL-1 Dysregulated expression or delayed expression associated, including diseases related to sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myeloid leukemia, insulin-dependent diabetes mellitus, atherosclerosis, nerve injury and senile degeneration. IL-1 And IL-1 Are structurally related polypeptides, at the amino acid level about 25% Homology of. Both are first synthesized into 31kDa Precursor, then cleaved to approximately 17.5kDa Of the mature protein. By Caspase-1/ICE Right IL-1 Cleavage of precursors is a critical step in the inflammatory response. Despite IL-1 And IL-1 Neither contain a typical hydrophobic signal peptide, but there is evidence that they can be secreted extracellularly through non-classical pathways. Unprocessed IL-1 Parts may be displayed on the cell membrane and may retain biological activity. With IL-1 The precursors are different, IL-1 Precursors vs. mature IL-1 Compared to having little or no biological activity. IL-1 Precursors vs. mature IL-1 All transported extracellularly. IL-1 And IL-1 Via immunoglobulin superfamily receptors and IL-1RA Combine to exert its effect. 80kDa Of I Type transmembrane receptor ( IL-1RI ) expressed in a variety of cells, including T Cells, fibroblasts, keratinocytes, endothelial cells, synovial lining cells, chondrocytes, and hepatocytes. 68kDa Of II Type transmembrane receptor ( IL-1RII ) in B Cells, neutrophils and bone marrow cells. IL-1RI And IL-1RII The extracellular region of 28% Homology of. But there are significant differences in other areas: II The intracellular domain of type receptor is only 29 Amino acids, and I The intracellular domain of type receptor is 213 Amino acids. IL-1RII It seems to be right IL-1 The signal does not respond, its function is to decoy the receptor and thus weaken IL-1 The role of. IL-1 Receptor ligand protein ( IL-1RAcP ) with IL-1RI The interaction is IL-1RI Necessary for signaling. IL-1RA Is a secreted molecule, is IL-1 Competitive inhibitors. Soluble IL-1RI And IL-1RII It is present in human plasma, joint fluid, and conditioned media of several cell lines. Moreover, vaccinia virus encodes similar to soluble IL-1RII Of IL-1 Binding proteins. |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| General Notes | 1. Please use the kit within the validity period. 2. The components of different kits and different batch kits cannot be mixed. 3. If the sample value is greater than the highest value of the standard curve, the sample should be diluted with diluent (1 ×) and re-tested; If the cell culture supernatant sample needs to be distributed and diluted, cell culture medium can be used for other intermediate dilutions except dilution with diluent in the last step. 4. Differences in test results can be caused by a variety of factors, including the operation of the experimenter, the use of the pipette, the plate washing technique, the reaction time or temperature, the storage of the kit, etc. 5. The terminating solution in the kit is an acidic solution. Please protect your glasses, hands, face and clothes when using it. 6. For scientific research only, not for in vitro diagnosis. |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Storage Temp. | Unopened kit, store at 2–8 °C | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Test Range | 62.5pg/mL-4000pg/mL |
Picture
Picture
Immunohistochemistry
