Frataxin ELISA Kit

Catalog No. ABIN7964493

Human Frataxin ELISA Kit, Colorimetric assay for quantification of Human Frataxin.

Quick Overview for Frataxin ELISA Kit (ABIN7964493)

Target: Frataxin (FXN)

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5 pg/mL - 4000 pg/mL

Application: ELISA

Sample Type: Biological Fluids, Plasma, Serum

Grade: Quick

Product Details Frataxin ELISA Kit

Minimum Detection Limit: 62.5 pg/mL

Purpose: Quick Human FXN (Frataxin) ELISA Kit

Analytical Method: Quantitative

Cross-Reactivity (Details): This kit recognizes Human FXN in samples. No significant cross-reactivity or interference between Human FXN and analogues was observed.

Sensitivity: 37.5 pg/mL

Components:

• Precoated Micro ELISA Plate, Dismountable
• Reference Standard
• Concentrated HRP Conjugate (100x)
• Reference Standard & Sample Diluent
• HRP Conjugate Diluent
• Concentrated Wash Buffer (25x)
• Substrate Reagent
• Stop Solution
• Plate Sealer
• Manual

Note: Reagents may contain slightly more volume than indicated on the label; mix and centrifuge the vial of reagents before use. Use precise measuring instruments rather than pouring directly from the vial.

Application Details

Sample Volume: 50 μL

Assay Time: 1.5 h

Plate: Pre-coated

Protocol:

1. Add 50 µL standard or sample to each well.
2. Add 50 µL HRP Conjugate. Incubate for 60 min at 37 °C.
3. Aspirate and wash 5 times.
4. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
5. Add 50 µL Stop Solution. Read at 450 nm immediately.
6. Calculation of results.

Reagent Preparation:

1. Bring all reagents to room temperature (18 - 25 °C) before use.
2. Standard Working Solution: Centrifuge the standard at 10,000xg for 1 min. Reconstitute with 1 mL of the Standard & Sample Diluent, allow to stand for 10 minutes, and gently invert several times until completely dissolved. Alternatively, vortex briefly at low speed, remove bubbles by low-speed centrifugation if necessary. This produces a 4000 pg/mL stock solution. Prepare serial dilutions according to experimental requirements. Recommended dilution gradient: 4000, 2000, 1000, 500, 250, 125, 62.5, 0 pg/mL. Dispense 500 μL of Standard & Sample Diluent into 7 EP tubes, respectively. Transfer 500 μL of the stock solution into the first tube. Sequentially transfer 500 μL from the preceding tube into the next tube for subsequent dilutions. Designate the last tube as the blank, do not transfer any solution into it. Serially diluted standards should be prepared immediately, prior to use.
3. HRP Conjugate Working Solution: Calculate the required amount before the experiment. When preparing, prepare slightly more than calculated. Centrifuge the Concentrated HRP Conjugate at 800×g for 1 min, then dilute the 100× Concentrated HRP Conjugate to 1× working solution with HRP Conjugate Diluent. Note: Prepare the working solution immediately, prior to use.
4. Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer. Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturation may occur in these samples, leading to false results. Samples should therefore be stored for a short period at 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thaw cycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged to remove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determine compatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibility of 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 the range of the standard curve, the optimal sample dilution for the particular experiment has to be determined.

Assay Precision:

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level of the target were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level of the target were tested on 3 different plates, 20 replicates in each plate.

Both intra-CV and inter-CV are < 10%.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment:

• For unopened kit: All reagents should be stored according to the labels on the vials. Protect Concentrated HRP Conjugate and Substrate Reagent from light. Always use the reconstituted standard within 24h. Reconstituted standard may be stored aliquoted at -20 °C for up to two weeks. Store samples at – 80 °C for up to 3 months. Avoid repeated freeze-thaw cycles.
• For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.

Expiry Date: 6 months

Target Details for Frataxin

Target: Frataxin (FXN)

Alternative Name: Frataxin intermediate form (i-FXN)

Background:

Target Synonym: Frataxin intermediate form (i-FXN)

Background: Frataxin protein (FXN) is a highly conserved mitochondrial protein. The mature form of Frataxin acts as a persulfide transferred to the scaffold protein ISCU as an activator, as a component of the core iron-sulfur cluster (ISC) assembly complex, and parti

Gene ID: 2395

UniProt: Q16595

Pathways: Transition Metal Ion Homeostasis, Regulation of Cell Size