Neurotrophin 4 ELISA Kit

Produktnummer ABIN1112671

Produktdetails Neurotrophin 4 ELISA Kit

Target: Neurotrophin 4 (NTF4)

Reaktivität: Ratte

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 600-20000 pg/mL

Untere Nachweisgrenze: 600 pg/mL

Applikation: ELISA

Verwendungszweck: For quantitative detection of NT-4 in rat serum, plasma, urine, cell culture supernatant or tissue samples.

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

Analytische Methode: Quantitative

Bestandteile: 1. One 96-well plate pre-coated with anti-rat NT-4 antibody 2. Standard: 0.5ml (22500pg /mL) 3. Standard diluent buffer: 1.5 ml 4. Wash buffer (30x): 20 ml.

Benötigtes Material: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Anwendungsinformationen

Kommentare:

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti-NT-4 antibody was pre-coated onto 96-well plates. And the HRP conjugated anti-NT-4 antibody was used as detection antibodies. The standards test samples and HRP conjugated detection antibody were added to the wells subsequently mixed and incubated then unbound conjugates were washed away with wash buffer. TMB substrates (A & B) were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the NT-4 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader and then the concentration of NT-4 can be calculated.

Plattentyp: Pre-coated

Aufbereitung der Reagenzien:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Aufbereitung der Proben:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles. Serum: Coagulate at room temperature for 10-20 °C min, then, centrifuge at the speed of 2000-3000 r.p.m. for 20 min to collect supernatant. If precipitation appeared, centrifuge again. Plasma: Collect plasma using EDTAor citrate plasma as an anticoagulant, and mix for 10-20 °C min, centrifuge at the speed of 2000-3000 r.p.m. for 20 min of collection. If precipitation appeared, centrifuge again. Urine: Collect urine using a sterile container, centrifuge at the speed of 2000-3000 r.p.m. for 20 min to collect supernatant. If precipitation appeared, centrifuge again. For collection of hydrothorax and cerebrospinal fluid, take reference to this operation. Cell culture supernatant: For secretory components: use a sterile container to collect. Centrifuge at the speed of 2000-3000 r.p.m. for 20 min to collect supernatant. For intracellular components: Dilute cell suspension with PBS(pH7.2-7.4) to make the cell concentration reached 1 million / ml. Damage cells and release of intracellular components through repeated freeze-thaw cycles. Centrifuge at the speed of 2000-3000 r.p.m. For 20 min to collect supernatant. If precipitation appeared, centrifuge again. Tissue samples: Cut samples and weight, add certain volume of PBS (pH7.4), rapidly frozen with liquid nitrogen. After melting, store samples at 2-8℃ . Add certain volume of PBS (pH7.4), homogenize thoroughly, centrifuge at the speed of 2000-3000 r.p.m. for 20 min to collect supernatant. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN 3 can not be used as test sample preservative, since it is the inhibitor for HRP. 3. After collecting samples, analyze immediately or aliquot and store frozen at -20 °C. Avoid repeated freeze-thaw cycles. 2. Wash buffer Dilute concentrated Wash buffer (Kit Component 4) 30-fold (1:30) with distilled water (i.e. add 20 ml of concentrated wash buffer into 580 ml of distilled water). 3. Standard Reconstitution of the Lyophilized Rat NT-4 standard (Kit Component 2): standard solution should be prepared no more than 2 hours prior to the experiment. Two tubes of standard are included in each kit. Use one tube for each experiment. (Note: Do not dilute the standard directly in the plate) a. 20,000pg/ml of standard solution: Add 0.5 ml of the 22,500pg/ml Standard (Kit Component 2) into 0.0625ml Standard diluent buffer (Kit Component 3) and mix thoroughly. b. 10,000 pg/ml -> 625 pg/ml of standard solutions: Label 5 Eppendorf tubes with 10,000 pg/ml, 5000 pg/ml, 2500 pg/ml, 1250 pg/ml, 625 pg/ml, respectively. Aliquot 0.2 ml of the Standard diluent buffer (Kit Component 3) into each tube. Add 0.2 ml of the above 20,000 pg/ml standard solution into 1st tube and mix thoroughly. Transfer 0.2 ml from 1st tube to 2nd tube and mix thoroughly. Transfer 0.2 ml from 2nd tube to 3rd tube and mix thoroughly, and so on. Chongqing Biospes Co., Ltd Product Manual

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Konservierungsmittel: Sodium azide, Thimerosal (Merthiolate)

Details zu Neurotrophin 4

Target: Neurotrophin 4 (NTF4)

Andere Bezeichnung: NT-4 (NTF4 Produkte)

Synonyme: NTF5 ELISA Kit, GLC10 ELISA Kit, GLC1O ELISA Kit, NT-4 ELISA Kit, NT-4/5 ELISA Kit, NT-5 ELISA Kit, NT4 ELISA Kit, NT5 ELISA Kit, NT4P ELISA Kit, Ntf5 ELISA Kit, 2900040K06Rik ELISA Kit, AI462899 ELISA Kit, NT4/5 ELISA Kit, Ntf-5 ELISA Kit, Ntf4 ELISA Kit, neurotrophin 4 ELISA Kit, neurotrophin 5 ELISA Kit, NTF4 ELISA Kit, Ntf4 ELISA Kit, ntf4 ELISA Kit, Ntf5 ELISA Kit

Hintergrund: Neurotrophins act as survival and differentiation factors in the nervous system and have been detected in the developing rodent testis. Three additional structurally homologous neurotrophic factors have been identified. These include brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4 (NT-4) (also designated NT-5). Neurotrophin-4 (NT-4), is a neurotrophic factor that signals predominantly through the TrkB receptor tyrosine kinase. Mammalian NT4 had many unusual features compared to the previously identified neurotrophins and was less conserved evolutionarily than the others, however, mammalian NT4 displayed bioactivity and TRK receptor specificity similar to that of Xenopus NT4.

Pathways: RTK Signalweg