HSP90AA1 ELISA Kit

Produktnummer ABIN579153

Produktdetails HSP90AA1 ELISA Kit

Target: HSP90AA1 (Heat Shock Protein 90kDa alpha (Cytosolic), Class A Member 1 (HSP90AA1))

Reaktivität: Ratte

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Applikation: ELISA

Verwendungszweck: This immunoassay kit allows for the specific measurement of rat HSP90 concentrations in serum and plasma.

Proben: Serum, Plasma

Analytische Methode: Quantitative

Spezifität: This assay recognizes recombinant and natural rat HSP90.

Kreuzreaktivität (Details): No significant cross-reactivity or interference was observed.

Produktmerkmale: Rattus norvegicus,Rat,Heat shock protein HSP 90-alpha,Heat shock 86 kDa,HSP 86,HSP86,Hsp90aa1,Hsp86,Hspca

Bestandteile: Reagent (Quantity ): Assay plate (1), Standard (2), Sample Diluent (1 × 20ml), Assay Diluent A (1x10ml), Assay Diluent B (1x10ml), Detection Reagent A (1 × 120μl), Detection Reagent B (1 × 120μl), Wash Buffer (25 x concentrate) (1 × 30ml), Substrate (1x10ml), Stop Solution (1 x 10ml)

Anwendungsinformationen

Probenmenge: 100 μL

Plattentyp: Pre-coated

Protokoll: This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for HSP90 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HSP90 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for HSP90 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of HSP90 bound in the initial step. The color development is stopped and the intensity of the color is measured.

Aufbereitung der Reagenzien:

Bring all reagents to room temperature before use. Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 500 mL of Wash Buffer. Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution of 10,000 pg/mL. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions. The undiluted standard serves as the high standard (10,000 pg/mL). The Sample Diluent serves as the zero standard (0 pg/mL). Detection Reagent A and B - Dilute to the working concentration specified on the vial label using Assay Diluent A and B (1:100), respectively.

Probennahme: Plasma - Fasting sample would be adaptive for this assay. Collect plasma using EDTA as an anticoagulant. Centrifuge samples for 15 minutes at 1000 x g at 2 - 8 °C within 30 minutes of collection. Store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles. Note: Citrate plasma has not been validated for use in this assay.

Testdurchführung:

Allow all reagents to reach room temperature. Arrange and label required number of strips.
1. Prepare all reagents, working standards and samples as directed in the previous sections.
2. Add 100 uL of Standard, Control, or sample* per well. Cover with the adhesive strip. Incubate for 2 hours at 37 °C.
3. Remove the liquid of each well, don’t wash.
4. Add 100 uL of Detection Reagent A to each well. Incubate for 1 hour at 37°C . Detection 3 Reagent A may appear cloudy. Warm to room temperature and mix gently until solution appears uniform.
5. Aspirate each well and wash, repeating the process three times for a total of three washes. Wash by filling each well with Wash Buffer (350 uL) using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
6. Add 100 uL of Detection Reagent B to each well. Cover with a new adhesive strip.Incubate for 1 hours at 37 °C.
7. Repeat the aspiration/wash as in step
5. 8. Add 90 uL of Substrate Solution to each well. Incubate for 30 minutes at room temperature. Protect from light.
9. Add 50 uL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
10. Determine the optical density of each well within 30 minutes, using a microplate reader set to 450 nm.
Important Note:1. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
2. It is recommended that no more than 32 wells be used for each assay run if manual pipetting is used since pipetting of all standards, specimens and controls should be completed within 5 minutes. A full plate of 96 wells may be used if automated pipetting is available.
3. Duplication of all standards and specimens, although not required, is recommended.
4. When mixing or reconstituting protein solutions, always avoid foaming.
5. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent.
6. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary.

Ergebnisberechnung:

Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the Ghrelin concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the 4 standard curve must be multiplied by the dilution factor.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: 1. The kit should not be used beyond the expiration date on the kit label.
2. Do not mix or substitute reagents with those from other lots or sources.
3. If samples generate values higher than the highest standard, further dilute the samples with the Assay Diluent and repeat the assay. Any variation in standard diluent, operator, pipetting technique, washing technique,incubation time or temperature, and kit age can cause variation in binding.
4. This assay is designed to eliminate interference by soluble receptors, ligands, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung: The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 °C upon being received. The other reagents can be stored at 4 °C.

Details zu HSP90AA1

Target: HSP90AA1 (Heat Shock Protein 90kDa alpha (Cytosolic), Class A Member 1 (HSP90AA1))

Andere Bezeichnung: Hsp90aa1 (HSP90AA1 Produkte)

Synonyme: EL52 ELISA Kit, HSP86 ELISA Kit, HSP89A ELISA Kit, HSP90A ELISA Kit, HSP90N ELISA Kit, HSPC1 ELISA Kit, HSPCA ELISA Kit, HSPCAL1 ELISA Kit, HSPCAL4 ELISA Kit, HSPN ELISA Kit, Hsp89 ELISA Kit, Hsp90 ELISA Kit, LAP2 ELISA Kit, Hsp86 ELISA Kit, Hspca ELISA Kit, htpG ELISA Kit, 86kDa ELISA Kit, 89kDa ELISA Kit, AL024080 ELISA Kit, AL024147 ELISA Kit, Hsp86-1 ELISA Kit, hsp4 ELISA Kit, HSP90 ELISA Kit, HSP90AA1 ELISA Kit, fb17b01 ELISA Kit, hsp90 ELISA Kit, hsp90a ELISA Kit, hsp90a.1 ELISA Kit, hsp90alpha ELISA Kit, wu:fb17b01 ELISA Kit, zgc:86652 ELISA Kit, Hsp90alpha ELISA Kit, heat shock protein 90 alpha family class A member 1 ELISA Kit, heat shock protein 90, alpha (cytosolic), class A member 1 ELISA Kit, Heat Shock Protein 90, cytosolic ELISA Kit, heat shock protein 90A ELISA Kit, molecular chaperone ELISA Kit, heat shock protein 90, alpha (cytosolic), class A member 1, tandem duplicate 1 ELISA Kit, heat shock protein HSP 90-alpha ELISA Kit, heat shock protein 90kDa alpha (cytosolic), class A member 1 ELISA Kit, HSP90AA1 ELISA Kit, Hsp90aa1 ELISA Kit, HSP90A ELISA Kit, hsp90A ELISA Kit, hsp90aa1.1 ELISA Kit, LOC108698781 ELISA Kit

Hintergrund: Heat shock proteins or HSPs are being synthesized under different kind of stress conditions and act as molecular chaperones for protein molecules. Because these proteins were first found in cells that were exposed to high temperatures， they are called ",heat shock proteins", and have been named according to their molecular weights. Usually HSPs are cytoplasmic proteins and they function in various intra-cellular processes. Heat shock proteins play an important role in protein-protein interactions， including folding and assisting in establishing of proper protein conformation， and prevention of inappropriate protein aggregation. HSP90 has been identified in the cytosol， nucleus and endoplasmic reticulum， and is reported to exist in many tissue types. In several tissues， including smooth muscle， HSP90 comprises up to 2% of total cellular protein. HSP90 functions as a dimer， assembled as part of heterocomplex. It possesses ATP-binding site and low ATPase activity. HSP90 is able to associate with actin filaments in certain conditions. Another important property of HSP90 is the binding of unoccupied steroid hormone receptors. HSP90 has been characterized as a molecular chaperone able to keep the target protein in a folding-competent state. It has an enhanced chaperone activity in oligomeric form at high temperatures. HSP90 function is sensitive to bivalent cations concentration.

Gen-ID: 3064

Pathways: M Phase, Regulation of Cell Size, Signaling Events mediated by VEGFR1 and VEGFR2, VEGFR1 Specific Signals