MED24 ELISA Kit

Catalog No. ABIN1770516

Product Details MED24 ELISA Kit

Target: MED24 (Mediator Complex Subunit 24 (MED24))

Reactivity: Monkey

Detection Method: Colorimetric

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma, Serum, Tissue Homogenate

Components:

• Microtiter plate (96 wells stripwell) - 1
• Enzyme conjugate - 1 vial
• Standard A - 1 vial
• Standard B - 1 vial
• Standard C - 1 vial
• Standard D - 1 vial
• Standard E - 1 vial
• Standard F - 1 vial
• Substrate A - 1 vial
• Substrate B - 1 vial
• Stop solution - 1 vial
• Wash solution - 1 vial
• Balance solution - 1 vial
• Instruction manual - 1

Material not included:

• Precision pipettors and disposable tips to deliver 10-1000 μL. A multi-channel pipette is desirable for large assays.
• 100 mL and 1 L graduated cylinders.
• Distilled or deionized water
• Tubes to prepare sample dilutions.
• Absorbent paper.
• Microplate reader capable of measuring absorbance at 450 nm.
• Centrifuge capable of 3000 x g.
• Microplate washer or washing bottle.
• Incubator (37 °C).
• Data analysis and graphing software.

Application Details

Application Notes:

• The supplier is only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of samples used in the whole test. Please reserve sufficient amounts of samples in advance.
• Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments.
• If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
• Owing to the possibility of mismatching between antigens from another resource and antibodies used in this supplier's kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by this supplier's products.
• Influenced by factors including cell viability, cell number and cell sampling time, samples from cell culture supernatant may not be recognized by the kit.
• Fresh samples without long time storage are recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.

Comment:

• It is recommended that all standards, controls and samples be run in duplicate. Standards and samples must be assayed at the same time.
• The coefficient of determination of the standard curve should be ≥ 0.95 and the highest O.D. should be more than 1.0.
• Cover or cap all kit components and store at 2-8 °C when not in use.
• Microtiter plates should be allowed to come to room temperature before opening the foil bags. Once the desired number of strips has been removed, immediately reseal the bag with desiccants and store at 2-8 °C to maintain plate integrity.
• Samples should be collected in pyrogen/endotoxin-free tubes.
• Samples should be frozen if not analyzed shortly after collection. Avoid multiple freeze-thaw cycles of frozen samples. Thaw completely and mix well prior to analysis.
• When possible, avoid use of badly hemolyzed or lipemic serum. If large amounts of particulate matter are present, centrifuge or filter prior to analysis.
• When pipetting reagents, maintain a consistent order of addition from well-to-well. This ensures equal incubation times for all wells.
• Do not mix or interchange different reagent lots from various kit lots.
• Do not use reagents after the kit expiration date.
• Read absorbance immediately after adding the stop solution.
• Incomplete washing will adversely affect the test outcome. All washing must be performed with Wash Solution provided. All residual wash liquid must be drained from the wells by efficient aspiration or by decantation followed by tapping the plate forcefully on absorbent paper. Never insert absorbent paper directly into the wells.
• Because TMB is light sensitive, avoid prolonged exposure to light. Also avoid contact between TMB and metal, otherwise color may develop.

Information on standard material:
Different kits have different standards. For kits detecting proteisn or peptidse, the standards are recombinant proteins or synthetic peptides. For kits detecting small chemical compounds, the standards are synthetic chemical compounds. There are no standards extracted from natural resources. All of our reombinant proteins are expressed in E.coli. The standard are dissolved in PBS with 0.1 % proclin 300 and some other preservatives.

Information on reagents:
The STOP solution is 1M sulphuric acid. The wash buffer is 0.05 % Tween 20 in PBS, pH 7.4. The ELISA kit dose not contain (sodium) azide, thimerosal, 2-mercaptoethanol (2-ME). Part of the reagents contain BSA.

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

Assay Time: 1.5 h

Plate: Pre-coated

Reagent Preparation:

• Samples - Please predict the concentration before assaying. If concentrations are unknown or not within the detection range, a preliminary experiment is recommended to determine the optimal dilution. PBS (pH 7.0-7.2) or 0.9 % physiological saline can be used as dilution buffer.
• Wash solution - Dilute 10 mL of wash solution concentrate (100×) with 990 mL of deionized or distilled water to prepare 1000 mL of wash solution (1×). If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have dissolved. The 1× wash solution is stable for 2 weeks at 2-8 °C.

Note:

• Bring all kit components and samples to room temperature (20-25 °C) before use.
• Do not dilute other ready-to-use components.

Sample Collection:

• Serum: Use a serum separator tube and allow samples to clot for 2 hours at room temperature or overnight at 2-8 °C. Centrifuge at approximately 1000 × g (or 3000 rpm) for 15 minutes. Remove serum and assay immediately or aliquot and store samples at -20 °C or -80 °C.
• Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 100 × g (or 3000 rpm) at 2-8 °C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20 °C or -80 °C.
• Tissue homogenates: The preparation of tissue homogenates will vary depending upon tissue type. For this assay, thoroughly rinse tissues in ice-cold PBS (0.02 mol/L, pH 7.0-7.2) to remove excess blood and weigh before homogenization. Mince the tissues into small pieces and homogenize them in a certain amount of PBS with a glass homogenizer on ice. Subject the resulting suspension to ultrasonication or to two freeze-thaw cycles to further break down cell membranes. After that, centrifuge for 15 minutes at 1500 × g (or 5000 rpm). Remove the supernate and assay immediately or aliquot and store samples at -20 °C or -80 °C.
• Cell lysates: Cells should be lysed according to the following directions.
  • 1. Adherent cells should be detached with trypsin and then collected by centrifugation. Suspension cells can be collected by centrifugation directly.
  • 2. Wash three times in PBS.
  • 3. Resuspend cells in PBS and subject to ultrasonication 3 times. Alternatively, freeze cells at -20 °C. Thaw cells with gentle mixing. Repeat the freeze/thaw cycle 3 times.
  • 4. Centrifuge at 1000 × g (or 3000 rpm) for 15 minutes at 2-8 °C to remove cellular debris.
  • 5. Assay immediately or store samples at -20 °C or -80 °C.
• Cell culture supernatants and other body fluids: Centrifuge cell culture media at 1000 × g (or 3000 rpm) for 15 minutes to remove debris. Assay immediately or store samples at -20 °C or -80 °C.

Note:

• Samples should be aliquoted and must be stored at -20 °C (≤3 months) or -80 °C (≤6 months) to avoid loss of bioactivity and contamination. If samples are to be run within 24 hours, they may be stored at 2-8 °C. Avoid repeated freeze-thaw cycles.
• Prior to assay, the frozen sample should be brought to room temperature slowly and mixed gently.
• Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts of certain chemicals.
• Samples containing a visible precipitate must be clarified prior to use in the assay. Care should be taken to minimize hemolysis. Do not use grossly hemolyzed or lipemic specimens.
• Do not use heat-treated specimens.

Restrictions: For Research Use only

Handling

Precaution of Use:

• This kit contains a small amount of 3, 3', 5, 5'-Tetramethylbenzidine (TMB) in Substrate B. TMB is non-carcinogenic but it is hazardous in case of skin contact, eye contact, ingestion and inhalation. In case of contact, rinse affected area with plenty of water.
• The stop solution provided with this kit is an acid solution. Wear protective gloves, clothing, and face protection.
• Care should be taken when handling the standard because of the known and unknown effects of it.
• Care should also be taken to avoid contact of skin or eyes with other kit reagents or specimens. In the case of contact, wash immediately with water.
• Do not pipette by mouth.
• Avoid generation of aerosols.
• Waste must be disposed of in accordance with federal, state and local environmental control regulations.
• All blood components and biological materials should be handled as potentially hazardous. Decontaminate and dispose specimens and all potentially contaminated materials as they could contain infectious agents. The preferred method of decontamination is autoclaving for a minimum of 1 hour a 121.5 °C.

Storage: 4 °C

Expiry Date: 6 months

Target Details for MED24

Target: MED24 (Mediator Complex Subunit 24 (MED24))

Alternative Name: Mediator of RNA polymerase II transcription subunit 24(MED24) (MED24 Products)

Synonyms: CG7999 ELISA Kit, Dmel\\CG7999 ELISA Kit, Med24 ELISA Kit, Trap100 ELISA Kit, dTRAP100 ELISA Kit, l(3)psg5 ELISA Kit, MED24 ELISA Kit, THRAP4 ELISA Kit, DpseGA20755 ELISA Kit, dpse_GLEANR_1506 ELISA Kit, GA20755 ELISA Kit, dyak_GLEANR_4540 ELISA Kit, DyakGE20715 ELISA Kit, GE20715 ELISA Kit, dmoj_GLEANR_17526 ELISA Kit, DmojGI16770 ELISA Kit, GI16770 ELISA Kit, dper_GLEANR_4776 ELISA Kit, DperGL22535 ELISA Kit, GL22535 ELISA Kit, dsim_GLEANR_14081 ELISA Kit, DsimGD14060 ELISA Kit, GD14060 ELISA Kit, ARC100 ELISA Kit, CRSP100 ELISA Kit, CRSP4 ELISA Kit, DRIP100 ELISA Kit, TRAP100 ELISA Kit, 911GSE ELISA Kit, AU040102 ELISA Kit, AW547152 ELISA Kit, D11Ertd307e ELISA Kit, Gse2 ELISA Kit, Pparb2 ELISA Kit, Pparbp2 ELISA Kit, R75526 ELISA Kit, Thrap4 ELISA Kit, fc05c04 ELISA Kit, thrap4 ELISA Kit, trap100 ELISA Kit, wu:fc05c04 ELISA Kit, Mediator complex subunit 24 ELISA Kit, mediator complex subunit 24 ELISA Kit, mediator of RNA polymerase II transcription subunit 24 ELISA Kit, GA20755 gene product from transcript GA20755-RA ELISA Kit, DyakGE20715-PA ELISA Kit, GI16770 gene product from transcript GI16770-RA ELISA Kit, GL22535 gene product from transcript GL22535-RA ELISA Kit, GD14060 gene product from transcript GD14060-RA ELISA Kit, microRNA 6884 ELISA Kit, MED24 ELISA Kit, LOC100016544 ELISA Kit, LOC100398369 ELISA Kit, Dpse\Trap100 ELISA Kit, DyakTrap100 ELISA Kit, Dmoj\Trap100 ELISA Kit, Dper\Trap100 ELISA Kit, Dsim\Trap100 ELISA Kit, Med24 ELISA Kit, MIR6884 ELISA Kit, med24 ELISA Kit

Pathways: Intracellular Steroid Hormone Receptor Signaling Pathway, Nuclear Hormone Receptor Binding, Stem Cell Maintenance, Regulation of Lipid Metabolism by PPARalpha, Positive Regulation of Endopeptidase Activity