MFGE8 ELISA Kit

Produktnummer ABIN6976240

Produktdetails MFGE8 ELISA Kit

Target: MFGE8 (Milk Fat Globule-EGF Factor 8 Protein (MFGE8))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 781.25 pg/mL - 50000 pg/mL

Untere Nachweisgrenze: 781.25 pg/mL

Applikation: ELISA

Verwendungszweck: For the quantitative determination of human milk fat globule-EGF factor 8 protein (MFGE8) concentrations in serum, cell culture supernates, urine and cerebrospinal fluid (CSF).

Proben: Cell Culture Supernatant, Cerebrospinal Fluid, Serum, Urine

Analytische Methode: Quantitative

Spezifität: This assay has high sensitivity and excellent specificity for detection of human MFGE8. No significant cross-reactivity or interference between human MFGE8 and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between human MFGE8 and all the analogues, therefore, cross reaction may still exist.

Sensitivität: 195.31 pg/mL

Bestandteile:

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

Anwendungsinformationen

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

Probenmenge: 100 μL

Testdauer: 1 - 4.5 h

Plattentyp: Pre-coated

Protokoll:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Aufbereitung der Reagenzien:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. 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).
4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 50000 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 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (50000 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.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• 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 Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Haltbarkeit: 6 months

Referenzen für MFGE8 ELISA Kit (ABIN6976240)

Zhang, Xie, Su, Li, Hu, Chen, Luo, Cui: "MFG-E8, a clearance glycoprotein of apoptotic cells, as a new marker of disease severity in chronic obstructive pulmonary disease." in: Brazilian journal of medical and biological research = Revista brasileira de pesquisas me?dicas e biolo?gicas / Sociedade Brasileira de Biofi?sica ... [et al.], Vol. 48, Issue 11, pp. 1032-8, (2015) (PubMed).

Yang, Hayashida, Forster, Li, Shen, Maheswaran, Chen, Anderson, Ellisen, Sgroi, Schmidt: "The integrin alpha(v)beta(3-5) ligand MFG-E8 is a p63/p73 target gene in triple-negative breast cancers but exhibits suppressive functions in ER(+) and erbB2(+) breast cancers." in: Cancer research, Vol. 71, Issue 3, pp. 937-45, (2011) (PubMed).

Details zu MFGE8

Target: MFGE8 (Milk Fat Globule-EGF Factor 8 Protein (MFGE8))

Andere Bezeichnung: milk fat globule-EGF factor 8 protein (MFGE8 Produkte)

Synonyme: BA46 ELISA Kit, EDIL1 ELISA Kit, HMFG ELISA Kit, HsT19888 ELISA Kit, MFG-E8 ELISA Kit, MFGM ELISA Kit, OAcGD3S ELISA Kit, SED1 ELISA Kit, SPAG10 ELISA Kit, hP47 ELISA Kit, ba46 ELISA Kit, edil1 ELISA Kit, oacgd3s ELISA Kit, MGC75581 ELISA Kit, hst19888 ELISA Kit, MGC145109 ELISA Kit, MFGE8 ELISA Kit, mfge8 ELISA Kit, zgc:154157 ELISA Kit, MGC99228 ELISA Kit, AA408458 ELISA Kit, AI325141 ELISA Kit, Mfgm ELISA Kit, P47 ELISA Kit, lactadherin ELISA Kit, AGS ELISA Kit, MFGME8 ELISA Kit, MFGMP-E8 ELISA Kit, im:6895406 ELISA Kit, mfge8l ELISA Kit, zgc:110846 ELISA Kit, milk fat globule-EGF factor 8 protein ELISA Kit, milk fat globule-EGF factor 8 protein b ELISA Kit, milk fat globule-EGF factor 8 protein L homeolog ELISA Kit, milk fat globule-EGF factor 8 protein a ELISA Kit, MFGE8 ELISA Kit, mfge8 ELISA Kit, mfge8b ELISA Kit, mfge8.L ELISA Kit, Mfge8 ELISA Kit, mfge8a ELISA Kit

Hintergrund:

Abbreviation: MFGE8

Alias: BA46, EDIL1, HsT19888, MFG-E8, OAcGD3S, SED1, SPAG10, hP47, O-acetyl disialoganglioside synthase|lactadherin|medin|sperm associated antigen 10|sperm surface protein hP47

UniProt: Q08431

Pathways: SARS-CoV-2 Protein Interaktom