TNF alpha ELISA Kit

Produktnummer ABIN8092020

Das Maus TNF alpha ELISA Kit (ABIN8092020) ist ein Colorimetric ELISA Kit zur Detektion von Maus TNF alpha.

Kurzübersicht für TNF alpha ELISA Kit (ABIN8092020)

Target: TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))

Reaktivität: Maus

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 1.563 pg/mL - 100 pg/mL

Applikation: ELISA

Proben: Cell Culture Supernatant, Cell Samples, Plasma, Serum, Tissue Lysate

Güteklasse: High Sensitivity

Produktdetails TNF alpha ELISA Kit

Untere Nachweisgrenze: 1.563 pg/mL

Verwendungszweck: Mouse TNF-α (High sensitive Tumor Necrosis Factor Alpha) high sensitivity ELISA Kit

Analytische Methode: Quantitative

Spezifität: Specifically binds with TNF-α , no obvious cross reaction with other analogues. Recommended Sample Dilution Ratio The following table shows the recommended dilution ratios for this kit for a limited number of samples for your reference only. (The matrix components in serum/plasma will affect the test results, which it need to be diluted at least 1/2 with Sample Dilution Buffer before testing! When the content of other samples is very low, the original solution can be added without dilution, but it is necessary to ensure that the pH is between 6.8 and 8.0, and it does not contain more than 10 % organic solvents or high-concentration protein denaturants.) 1. Common sample validation: Sample Type Recommended Dilution Ratio Content Normal mouse serum (n=16) 1/2 dilution ND-25pg/mL Normal mouse plasma (EDTA, Citrate , heparin) (n=16) 1/2 dilution ND-31pg/mL RAW 264.7 cells were cultured with 5 %FBS + 1640 + double antibody for 12 hours 1/2dilution 28pg/mL RAW 264.7 cells were cultured with 5 %FBS+ 1640 + double antibody +1 μg/mL LPS, and the cell culture supernatant was detected after 12 hours 1/5-1/20 dilution 618pg/mL RAW 264.7 cells were cultured with 5 %FBS+ 1640 + double antibody +1 μg/mL LPS for 12 hours. After that, 300 ng/mL Brefeldin A (BFA) was added and cultured for 3 hours. Add cell lysis buffer(Catalogue No.:E050), collect the lysate solution (total protein concentration measured by BCA assay: 2.39 mg/mL) to detect. 1/50 dilution 1.31 ng/mg(total protein) 2. KD sample validation (Detect TNF-α-KD RAW 264.7 cells): Sample Type Dilution Ratio Content Wild RAW 264.7 cells were stimulated with 1 μg/mL LPS for 12 hours to detect cell culture supernatant 1/20 dilution 711pg/mL RAW 264.7 cells (+siRNA) were stimulated with 1 μg/mL LPS for 12 hours to detect cell culture supernatants 1/2 dilution ND RAW 264.7 cells (+siRNA) were treated with 1 μg/mL LPS for 12 hours and then cultured with 300 ng/mL Brefeldin A (BFA) for 3 hours. Add cell lysis buffer(Catalogue No.:E050), collect the lysate solution (total protein concentration measured by BCA assay: 1.75 mg/mL) to detect. 1/2 dilution 17pg/mg(total protein) Note:ND is lower than the sensitivity of the kit and was not detected 3. Antibody by WB KD validation (Detect TNF-α-KO RAW 264.7 cells):

Sensitivität: 0.938 pg/mL

Bestandteile:

• ELISA Microplate(Dismountable)
• Lyophilized Standard
• Biotin-labeled Antibody
• HRP-Streptavidin Conjugate(SABC
• 100X)
• TMB Substrate
• Sample Dilution Buffer
• Stop Solution
• Plate Sealer
• Product Description

Benötigtes Material:

• Microplate reader (wavelength: 450nm)
• 37 °C incubator (CO2 incubator for cell culture is not recommenced.)
• Automated plate washer or multi-channel pipette/5 mL pipettor
• Precision single (0.5-10μL
• 5-50μL
• 20-200μL
• 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
• Sterile tubes and Eppendorf tubes with disposable tips
• Absorbent paper and loading slot
• Deionized or distilled water

Anwendungsinformationen

Applikationshinweise: Optimal working dilution should be determined by the investigator.

Kommentare:

Sandwich ELISA, Double Antibody

Probenmenge: 100 μL

Testdauer: 4 h

Plattentyp: Pre-coated

Protokoll: Step 1: Add 100 µL standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37 °C. Washing: Wash the plate twice without immersing. Step 2: Add 100 µL biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37 °C. Washing: Wash the plate three times and immerse for 1 min each time. Step 3: Add 100 µL HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37 °C. Washing: Wash the plate five times and immerse for 1 min each time. Step 4: Add 90 µL TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37 °C. (Accurate TMB visualization control is required.) Step 5: Add 50 µL stop solution. Read at 450nm immediately and calculate.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: Avoid direct light exposure to antibody solutions

Lagerung: 4 °C

Informationen zur Lagerung: Store sealed Kit at 2-8° C. Do Not Cryopreserve. Store ELISA Microplate for 1 month at 2-8°C, Store for 12 month at -20°C. Put the leftover strips into a sealed foil bag with the desiccant

Haltbarkeit: 12 months

Details zu TNF alpha

Target: TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))

Andere Bezeichnung: TNF-alpha

Hintergrund: Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a, Tumor necrosis factor, membrane form, N-terminal fragment, NTF, Intracellular domain 1, ICD1, Intracellular domain 2, ICD2, C-domain 1, C-domain 2, Tumor necrosis factor, soluble form, TNF, TNFA, TNFSF2

UniProt: P06804

Pathways: NF-kappaB Signalweg, Apoptose, Caspase Kaskade in der Apoptose, TLR Signalweg, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus, Inflammasome