NFkB ELISA Kit

Produktnummer ABIN6958236

Produktdetails NFkB ELISA Kit

Target: NFkB (Nuclear Factor kappa B (NFkB))

Reaktivität: Ratte

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 0.31 ng/mL - 20 ng/mL

Untere Nachweisgrenze: 0.31 ng/mL

Applikation: ELISA

Verwendungszweck: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of NFkB in rat tissue homogenates, cell lysates, cell culture supernates.

Proben: Cell Culture Supernatant, Cell Lysate, Tissue Homogenate

Analytische Methode: Quantitative

Spezifität: This assay has high sensitivity and excellent specificity for detection of Nuclear Factor Kappa B (NFkB)

Sensitivität: 0.114 ng/mL

Bestandteile:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Anwendungsinformationen

Kommentare:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Probenmenge: 100 μL

Testdauer: 3 h

Plattentyp: Pre-coated

Protokoll:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Aufbereitung der Reagenzien:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 40 ng/mL. Firstly dilute the stock solution to 20 ng/mL and the diluted standard serves as the highest standard (20 ng/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. 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): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Vorsichtsmaßnahmen: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Haltbarkeit: 6 months

Referenzen für NFkB ELISA Kit (ABIN6958236)

Su, Yu, Wang, Peng, Bai, Li, Li, Xia, Fu, Liu, Liu, Kang: "Na+/K+-ATPase Alpha 2 Isoform Elicits Rac1-Dependent Oxidative Stress and TLR4-Induced Inflammation in the Hypothalamic Paraventricular Nucleus in High Salt-Induced Hypertension." in: Antioxidants (Basel, Switzerland), Vol. 11, Issue 2, (2022) (PubMed).

Lv, Li, Hu, Wang, Wu, Li et al.: "The shear wave elastic modulus and the increased nuclear factor kappa B (NF-kB/p65) and cyclooxygenase-2 (COX-2) expression in the area of myofascial trigger points activated in a rat model by blunt ..." in: Journal of biomechanics, Vol. 66, pp. 44-50, (2019) (PubMed).

George, Esmat, Tadros, El-Demerdash: "In vivo cellular and molecular gastroprotective mechanisms of chrysin; Emphasis on oxidative stress, inflammation and angiogenesis." in: European journal of pharmacology, Vol. 818, pp. 486-498, (2018) (PubMed).

Baluchnejadmojarad, Zeinali, Roghani: "Scutellarin alleviates lipopolysaccharide-induced cognitive deficits in the rat: Insights into underlying mechanisms." in: International immunopharmacology, Vol. 54, pp. 311-319, (2018) (PubMed).

Khajevand-Khazaei, Ziaee, Motevalizadeh, Rohani, Afshin-Majd, Baluchnejadmojarad, Roghani: "Naringenin ameliorates learning and memory impairment following systemic lipopolysaccharide challenge in the rat." in: European journal of pharmacology, Vol. 826, pp. 114-122, (2018) (PubMed).

Fayed, El-Naga, Akool, El-Demerdash: "The potential antifibrotic impact of apocynin and alpha-lipoic acid in concanavalin A-induced liver fibrosis in rats: Role of NADPH oxidases 1 and 4." in: Drug discoveries & therapeutics, Vol. 12, Issue 2, pp. 58-67, (2018) (PubMed).

Afshin-Majd, Bashiri, Kiasalari, Baluchnejadmojarad, Sedaghat, Roghani: "Acetyl-l-carnitine protects dopaminergic nigrostriatal pathway in 6-hydroxydopamine-induced model of Parkinson's disease in the rat." in: Biomedicine & pharmacotherapy, Vol. 89, pp. 1-9, (2017) (PubMed).

Kiasalari, Heydarifard, Khalili, Afshin-Majd, Baluchnejadmojarad, Zahedi, Sanaierad, Roghani: "Ellagic acid ameliorates learning and memory deficits in a rat model of Alzheimer's disease: an exploration of underlying mechanisms." in: Psychopharmacology, Vol. 234, Issue 12, pp. 1841-1852, (2017) (PubMed).

Sedaghat, Taab, Kiasalari, Afshin-Majd, Baluchnejadmojarad, Roghani: "Berberine ameliorates intrahippocampal kainate-induced status epilepticus and consequent epileptogenic process in the rat: Underlying mechanisms." in: Biomedicine & pharmacotherapy, Vol. 87, pp. 200-208, (2017) (PubMed).

Diling, Tianqiao, Jian, Chaoqun, Ou, Yizhen: "Docking Studies and Biological Evaluation of a Potential β-Secretase Inhibitor of 3-Hydroxyhericenone F from Hericium erinaceus." in: Frontiers in pharmacology, Vol. 8, pp. 219, (2017) (PubMed).

Diling, Chaoqun, Jian, Jian, Jiyan, Yizhen, Guoxiao: "Immunomodulatory Activities of a Fungal Protein Extracted from Hericium erinaceus through Regulating the Gut Microbiota." in: Frontiers in immunology, Vol. 8, pp. 666, (2017) (PubMed).

Abd El Motteleb, Ibrahim, Elshazly: "Sildenafil protects against bile duct ligation induced hepatic fibrosis in rats: Potential role for silent information regulator 1 (SIRT1)." in: Toxicology and applied pharmacology, Vol. 335, pp. 64-71, (2017) (PubMed).

Kiasalari, Khalili, Baluchnejadmojarad, Roghani: "Protective Effect of Oral Hesperetin Against Unilateral Striatal 6-Hydroxydopamine Damage in the Rat." in: Neurochemical research, Vol. 41, Issue 5, pp. 1065-72, (2016) (PubMed).

El-Agroudy, El-Naga, El-Razeq, El-Demerdash: "Forskolin, a hedgehog signalling inhibitor, attenuates carbon tetrachloride-induced liver fibrosis in rats." in: British journal of pharmacology, Vol. 173, Issue 22, pp. 3248-3260, (2016) (PubMed).

Kiasalari, Rahmani, Mahmoudi, Baluchnejadmojarad, Roghani: "Diosgenin ameliorates development of neuropathic pain in diabetic rats: Involvement of oxidative stress and inflammation." in: Biomedicine & pharmacotherapy, Vol. 86, pp. 654-661, (2016) (PubMed).

Ahshin-Majd, Zamani, Kiamari, Kiasalari, Baluchnejadmojarad, Roghani: "Carnosine ameliorates cognitive deficits in streptozotocin-induced diabetic rats: Possible involved mechanisms." in: Peptides, Vol. 86, pp. 102-111, (2016) (PubMed).

Baluchnejadmojarad, Kiasalari, Afshin-Majd, Ghasemi, Roghani: "S-allyl cysteine ameliorates cognitive deficits in streptozotocin-diabetic rats via suppression of oxidative stress, inflammation, and acetylcholinesterase." in: European journal of pharmacology, Vol. 794, pp. 69-76, (2016) (PubMed).

Zarezadeh, Baluchnejadmojarad, Kiasalari, Afshin-Majd, Roghani: "Garlic active constituent s-allyl cysteine protects against lipopolysaccharide-induced cognitive deficits in the rat: Possible involved mechanisms." in: European journal of pharmacology, Vol. 795, pp. 13-21, (2016) (PubMed).

Hijová, Šoltésová, Salaj, Kuzma, Strojný, Bomba, Gregová: "Preventive use of Lactobacillus plantarum LS/07 and inulin to relieve symptoms of acute colitis." in: Acta biochimica Polonica, Vol. 62, Issue 3, pp. 553-7, (2015) (PubMed).

Ragab, Abdallah, El-Abhar: "Cilostazol renoprotective effect: modulation of PPAR-?, NGAL, KIM-1 and IL-18 underlies its novel effect in a model of ischemia-reperfusion." in: PLoS ONE, Vol. 9, Issue 5, pp. e95313, (2014) (PubMed).

Details zu NFkB

Target: NFkB (Nuclear Factor kappa B (NFkB))

Abstract: NFkB Produkte

Synonyme: EBP-1 ELISA Kit, KBF1 ELISA Kit, NF-kB1 ELISA Kit, NF-kappa-B ELISA Kit, NF-kappaB ELISA Kit, NFKB-p105 ELISA Kit, NFKB-p50 ELISA Kit, NFkappaB ELISA Kit, p105 ELISA Kit, p50 ELISA Kit, NF-kB ELISA Kit, NF-KB1 ELISA Kit, NF-kappaB1 ELISA Kit, p50/p105 ELISA Kit, nuclear factor kappa B subunit 1 ELISA Kit, nuclear factor of kappa light polypeptide gene enhancer in B cells 1, p105 ELISA Kit, NFKB1 ELISA Kit, Nfkb1 ELISA Kit

Pathways: Ubiquitin Proteasome Pathway, S100 Proteine