RASA1 ELISA Kit

Catalog No. ABIN6229251

This Colorimetric ELISA kit is designed for the quantitative measurement of Human RASA1.

Quick Overview for RASA1 ELISA Kit (ABIN6229251)

Target: RASA1 (RAS P21 Protein Activator (GTPase Activating Protein) 1 (RASA1))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.156 ng/mL - 10 ng/mL

Application: ELISA

Sample Type: Cell Lysate, Tissue Homogenate

Product Details RASA1 ELISA Kit

Minimum Detection Limit: 0.156 ng/mL

Purpose: This kit is based on sandwich enzyme-linked immuno-sorbent assay technology.

Analytical Method: Quantitative

Sensitivity: < 0.06 ng/mL

Components: The kit components listed are for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• Pre-coated 96-Well Microplate
• Standard
• Standard Diluent Buffer
• Wash Buffer
• Detection Reagent A
• Detection Reagent B
• Diluent A
• Diluent B
• TMB Substrate
• Stop Solution
• Plate Sealer

Material not included:

• 37 °C incubator
• Multi and single channel pipettes and sterile pipette tips
• Squirt bottle or automated microplate washer
• 1.5 mL tubes
• Distilled water
• Absorbent filter papers
• 100 mL and 1 liter graduated cylinders
• Microplate reader (wavelength: 450 nm)
• ELISA Shaker

Application Details

Plate: Pre-coated

Protocol: This kit is based on sandwich enzyme-linked immuno-sorbent assay technology. An antibody is pre-coated onto a 96-well plate. Standards, test samples, and biotin-conjugated reagent are added to the wells and incubated. The HRP-conjugated reagent is then added, and the whole plate is incubated. Unbound conjugates are removed using wash buffer at each stage. TMB substrate is used to quantify the HRP enzymatic reaction. After TMB substrate is added, only wells that contain sufficient GSTa2 will produce a blue coloured product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow colour is proportional to the GSTa2 amount bound on the plate. The Optical Density (OD) is measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of GSTa2 can be calculated.

Reagent Preparation:

This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• 1) Standard: Prepare the standard with the recommended volume of Standard Diluent Buffer, to make the standard solution. Then use the Standard Diluent buffer to carry out serial dilutions of the standard solution, as instructed in the Protocol.

• 2) Wash Buffer: Dilute the concentrated Wash Buffer with distilled water, as instructed in the Protocol.

• 3) Detection Reagent Preparation: Calculate the total volume of working solution required. Dilute Detection Reagent A and Detection Reagent B with Diluent A and Diluent B, respectively, at 1:100.

Sample Collection:

• Serum: Samples should be collected into a serum separator tube. Coagulate the serum by leaving the tube undisturbed in a vertical position overnight at 4 °C or at room temperature for up to 60 minutes. Centrifuge at approximately 1000 x g for 20 min. Analyze the serum immediately or aliquot and store at -20 °C or -80 °C.

• Plasma: Collect plasma using heparin or EDTA as an anticoagulant. Centrifuge for 15 minutes at 1000 x g within 30 minutes of collection. Assay immediately or aliquot and store at -20 °C or -80 °C. Avoid hemolysis and high cholesterol samples.

• Tissue homogenates: The preparation of tissue homogenates will vary depending upon tissue type - this is just an example. Rinse tissues with ice-cold PBS to remove the excess of blood. Weigh before homogenization. Finely mince tissues and homogenize with a tissue homogenizer on ice in PBS and sonicate the cell suspension. Centrifuge the homogenates at 5000 x g for 5 min and collect the supernatant. Assay immediately or aliquot and store at -20 °C.

Assay Procedure:

This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• 1) Set standard, test samples and control wells.

• 2) Aliquot 100 μL of diluted standard into the standard wells.

• 3) Aliquot 100 μL of Standard Diluent buffer into control (zero) well.

• 4) Aliquot 100 μL of diluted samples into the sample wells. Incubate for 1 hr at 37 °C.

• 5) Aliquot 100 μL of Detection Reagent A to each well. Incubate for 1 hr at 37 °C.

• 6) Wash 3 times.

• 7) Aliquot 100 μL of Detection Reagent B to each well. Incubate for 90 mins at 37 °C.

• 8) Wash 5 times.

• 9) Aliquot 90 μL of TMB Substrate to each well. Incubate for 10-20 mins at 37 °C.

• 10) Aliquot 50 μL of Stop Solution.

• 11) Measure the OD at 450 nm.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: Shipped at 4°C. Upon receipt, store the kit according to the storage instruction in the kit's manual.The stability of the kit is determined by the rate of activity loss. The loss rate is less than 5% within the expiration date under appropriate storage conditions. To minimize performance fluctuations, operation procedures and lab conditions should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same user throughout.

Expiry Date: 6 months

Target Details for RASA1

Target: RASA1 (RAS P21 Protein Activator (GTPase Activating Protein) 1 (RASA1))

Alternative Name: Ras GTPase-activating protein 1

Gene ID: 5921

HGNC: 9871

OMIM: 139150

UniProt: P20936

Pathways: Regulation of Actin Filament Polymerization, Signaling of Hepatocyte Growth Factor Receptor, VEGFR1 Specific Signals