IL-1 beta ELISA Kit

Produktnummer ABIN625201

Produktdetails IL-1 beta ELISA Kit

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Reaktivität: Ratte

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Applikation: ELISA

Verwendungszweck: Rat IL-1 beta (IL-1 F2) ELISA Kit for cell and tissue lysate samples.

Proben: Cell Lysate, Tissue Lysate

Analytische Methode: Quantitative

Spezifität: The antibody pair provided in this kit recognizes rat IL-1 beta.

Kreuzreaktivität (Details): This ELISA kit shows no cross-reactivity with any of the following cytokines tested: rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1alpha, IL-1beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3alpha, beta- NGF, TIMP-1, VEGF.

Sensitivität: 80 pg/mL

Produktmerkmale:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Bestandteile:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Benötigtes Material:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis
• Cell lysate buffer

Anwendungsinformationen

Probenmenge: 100 μL

Plattentyp: Pre-coated

Protokoll:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Aufbereitung der Reagenzien:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
   2. Sample dilution: Tissue lysate and cell lysate sample should be diluted at least 5-fold with 1x Sample Diluent Buffer.
   3. Sample Diluent Buffer (Item D) and Assay Diluent (Item E) should be diluted 5-fold with deionized or distilled water before use.
   4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL 1x Sample Diluent Buffer (Item D, should be diluted 5-fold with deionized or distilled water before use) into Item C vial to prepare a 50,000 pg/mL standard. Dissolve the powder thoroughly by a gentle mix. Pipette 260 µL 1x Sample Diluent Buffer into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Sample Diluent Buffer serves as the zero standard (0 pg/mL). Standard vial + 400 µL 130 µL 130myl 130 µL 130 µL 130 µL 130 µL 50,000 16,667 5556 1852 617.3 205.8 68.59 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
   5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
   6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diuent into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diuent and used in step 4 of Part VI Assay Procedure.
   7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) before use. HRP-Streptavidin concentrate should be diluted 200-fold with 1x Assay Diuent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 50 µL of HRP-Streptavidin concentrate into a tube with 10 ml 1x Assay Diluent to prepare a 200-fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.
   8. Cell lysate buffer should be diluted 2-fold with deionized or distilled water (for cell lysate and tissue lysate).

Testdurchführung:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
   2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
   3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
   4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
   5. Discard the solution. Repeat the wash as in step
   6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
   7. Discard the solution. Repeat the wash as in step
   8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
   9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Ergebnisberechnung:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Sample Diluent Buffer Rat IL-1beta concentration (pg/mL) O D =4 50 (n m ) 0.001 0.01 0.1 1 10 10 100 1,000 10,000 100,000
Sensitivity: The minimum detectable dose of IL-1beta is typically less than 80 pg/mL.
Recovery: Recovery was determined by spiking various levels of Rat IL-1beta into Rat tissue lysate and cell lysate. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Tissue lysate 136.5 126-145 Cell lysate 96.22 88-105
Linearity: Sample Type Tissue Cell Lysate lysate 1:2 Average % of 110.8 134.2 Expected Range ( %) 99-118 123-139 1:4 Average % of 135.4 118.5 Expected Range ( %) 125-142 109-127
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Testpräzision: Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: Avoid repeated freeze-thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Haltbarkeit: 6 months

Referenzen für IL-1 beta ELISA Kit (ABIN625201)

Bijjem, Padi, lal Sharma: "Pharmacological activation of heme oxygenase (HO)-1/carbon monoxide pathway prevents the development of peripheral neuropathic pain in Wistar rats." in: Naunyn-Schmiedeberg's archives of pharmacology, Vol. 386, Issue 1, pp. 79-90, (2013) (PubMed).

Kumar, Gupta, Srinivasan, Nag, Srivastava, Saxena, Jha: "Hesperetin rescues retinal oxidative stress, neuroinflammation and apoptosis in diabetic rats." in: Microvascular research, Vol. 87, pp. 65-74, (2013) (PubMed).

Details zu IL-1 beta

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Andere Bezeichnung: IL-1 beta (IL1B Produkte)

Synonyme: IL-1 ELISA Kit, IL1-BETA ELISA Kit, IL1F2 ELISA Kit, IL-1BETA ELISA Kit, IL1beta ELISA Kit, il1-b ELISA Kit, zgc:111873 ELISA Kit, IL-1B ELISA Kit, IL-1beta ELISA Kit, Il-1b ELISA Kit, IL1B ELISA Kit, IL-1 beta ELISA Kit, IL-1b ELISA Kit, interleukin 1 beta ELISA Kit, interleukin 1, beta ELISA Kit, IL1B ELISA Kit, il1b ELISA Kit, Il1b ELISA Kit

Hintergrund: Interleukin-1 beta (IL-1 beta)

UniProt: Q63264

Pathways: NF-kappaB Signalweg, Interferon-gamma Pathway, TLR Signalweg, Negative Regulation of Hormone Secretion, Cellular Response to Molecule of Bacterial Origin, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Myometrial Relaxation and Contraction, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Autophagie, Cancer Immune Checkpoints, Inflammasome