TNF alpha ELISA Kit

Produktnummer ABIN6973625

Produktdetails TNF alpha ELISA Kit

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

Reaktivität: Huhn

Nachweismethode: Colorimetric

Methodentyp: Competition ELISA

Detektionsbereich: 0.27 pg/mL - 200 pg/mL

Untere Nachweisgrenze: 0.27 pg/mL

Applikation: ELISA

Verwendungszweck: For the quantitative determination of chicken tumor necrosis factor α (TNF-α) concentrations in serum, plasma, tissue homogenates.

Proben: Plasma, Serum, Tissue Homogenate

Analytische Methode: Quantitative

Spezifität: This assay has high sensitivity and excellent specificity for detection of chicken TNF-α. No significant cross-reactivity or interference between chicken TNF-α and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between chicken TNF-α and all the analogues, therefore, cross reaction may still exist.

Bestandteile:

• Assay plate
• Standard
• HRP-conjugate (100 x concentrate)
• Sample Diluent
• HRP-conjugate Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Stop Solution

Adhesive Strip

Anwendungsinformationen

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

Probenmenge: 50 μL

Testdauer: 1 - 4.5 h

Plattentyp: Pre-coated

Protokoll:

1. Prepare reagents, samples and standards as instructed.
2. Set a Blank well without any solution.
3. Add 50 µL standard or sample to each well.
4. Add 50 µL HRP-conjugate (1x) to each well (Not to Blank well).
5. Incubate 1 hour at 37 °C
6. Aspirate and wash 5 times.
7. Add 90 μL of TMB Substrate to each well. Incubate for 20 minutes at 37 °C. Protect from light.
8. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Aufbereitung der Reagenzien:

1. HRP-conjugate (1x) - Centrifuge the vial before opening. HRP-conjugate requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-conjugate + 990 μL of HRP-conjugate Diluent.
2. Wash Buffer(1x)- If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
3. Standard Centrifuge the standard vial at 6000-10000rpm for 30s before opening. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 200 pg/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 200 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 3-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (200 pg/mL). Sample Diluent serves as the zero standard (0 pg/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Distilled water is recommended to be used to make the preparation for reagents. Contaminated water or container for reagent preparation will influence the detection result.

Aufbereitung der Proben:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Testpräzision: Intra-assay Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date. May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If don't make recent use, better keep it store at HRP-conjugate -20°C. Opened kit HRP-conjugate Diluent Sample Diluent May be stored for up to 1 month at 2 - 8°C. Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Haltbarkeit: 6 months

Referenzen für TNF alpha ELISA Kit (ABIN6973625)

Alhenaky, Abdelqader, Abuajamieh, Al-Fataftah: "The effect of heat stress on intestinal integrity and Salmonella invasion in broiler birds." in: Journal of thermal biology, Vol. 70, Issue Pt B, pp. 9-14, (2018) (PubMed).

Wang, Zeng, Wang, Zhu, Peng, Hou, Thacker, Qiao: "The antimicrobial peptide sublancin ameliorates necrotic enteritis induced by Clostridium perfringens in broilers." in: Journal of animal science, Vol. 93, Issue 10, pp. 4750-60, (2015) (PubMed).

Zhong, Shi, Chen, Xu, Wang, Beier, Hou, Liu: "Polyphenol extracts from Punica granatum and Terminalia chebula are anti-inflammatory and increase the survival rate of chickens challenged with Escherichia coli." in: Biological & pharmaceutical bulletin, Vol. 37, Issue 10, pp. 1575-82, (2014) (PubMed).

Details zu TNF alpha

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

Andere Bezeichnung: Tumor necrosis factor alpha,TNF-alpha (TNF alpha Produkte)

Synonyme: DIF ELISA Kit, TNF-alpha ELISA Kit, TNFA ELISA Kit, TNFSF2 ELISA Kit, RATTNF ELISA Kit, Tnfa ELISA Kit, tnf ELISA Kit, TNF-a ELISA Kit, TNFalpha ELISA Kit, Tnfsf1a ELISA Kit, TNFa ELISA Kit, cTNF ELISA Kit, Tnf-alpha ELISA Kit, tnfa-like ELISA Kit, TNF-ALPHA ELISA Kit, dif ELISA Kit, tnfa ELISA Kit, xtnf ELISA Kit, tnfsf2 ELISA Kit, tnf-alpha ELISA Kit, Cachectin ELISA Kit, tumor necrosis factor ELISA Kit, tumor necrosis factor b (TNF superfamily, member 2) ELISA Kit, tumor necrosis factor alpha ELISA Kit, tumor necrosis factor a (TNF superfamily, member 2) ELISA Kit, TNF ELISA Kit, Tnf ELISA Kit, tnf ELISA Kit, tnfb ELISA Kit, tnf-alpha ELISA Kit, LOC103694380 ELISA Kit, tnfa ELISA Kit

Hintergrund:

Abbreviation: TNF-α

Alias: DADB-70P7.1, DIF, TNF-alpha, TNFA, TNFSF2, APC1 protein|TNF superfamily, member 2|TNF, macrophage-derived|TNF, monocyte-derived|cachectin|tumor necrosis factor alpha

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