Hypersensitive ECL Chemiluminescence Kit（Femtogram） servicebio G2020-500ML
$50.00
In stock
SKU
G2020-500ML
Product Information
| Product Name: Hypersensitive ECL Chemiluminescence Kit（Femtogram） | Cat. No.: G2020- 5 0 ML |
| Spec.: 2×25 mL | G2020- 1 00 ML |
| 2× 5 0 mL | G2020- 5 00 ML |
| 2× 25 0 mL |
Product Description/Introduction
ECL chemiluminescence Kit is a highly sensitive chemiluminescence kit based on luminol ECL chemiluminescence. This product can react with horseradish peroxidase (HRP) conjugated on secondary antibody to produce fluorescence and detect samples by using X-ray lithography or other appropriate fluorescence imaging equipment (fluorescence or chemiluminescence imaging system, etc.). Low concentration samples can still be detected, saving samples. Signal can still be detected after incubation with low concentrations of antibodies (primary, secondary), saving precious antibodies. Signal values are enhanced at least 10-fold compared to the Sensitive ECL Chemiluminescence Kit (G2074).
Storage and Shipping Conditions
Ship with wet ice; Store in the dark at 2-8℃, valid for 12 months.
Product Components
| Component Number: G2020-1 | Component: Hypersensitive ECL solution A |
| G2020-50ML: 25 mL | G 2020- 10 0ML: 50 mL |
| G 2020-5 0 0ML: 250 mL | Component Number: G2020-2 |
| Component: Hypersensitive ECL solution B | G2020-50ML: 25 mL |
| G 2020- 10 0ML: 50 mL | G 2020-5 0 0ML: 250 mL |
| Product Manual | 1 pc |
Assay Protocol/Procedures
1. Preparation of hypersensitive ECL working solution: Mix the hypersensitive ECL solution A and hypersensitive ECL solution B in equal volumes, room temperature preparation, ready to use.
2. In Western experiment, PVDF membrane (or NC membrane) is incubated with secondary antibody, washed several times, and excess liquid is absorbed by filter paper. Stick two layers of PE gloves or other transparent films on the exposure box, place the protein side of the PVDF film between the two layers of the exposure box, add the mixed ECL working solution to cover the film and place it on the film for 1-2 min.
3. Remove the ECL working liquid with filter paper or absorbent paper, cover the upper film and start pressing the film.
4. The pressed film should be developed and fixed with developing and fixing reagents (G2019, G2023, G2024 are recommended). Adjust exposure conditions according to luminous intensity.
Note
1. Be sure to change the pipette tips during pipetting of Reagent A and B to avoid cross-contamination, which may result in the failure of active components.
2. Please wear gloves and use clean equipment such as tweezers to avoid contamination by exogenous proteins and metal ions during contact with the membrane.
3. Sodium azide inhibits HRP activity and all related reagents should be avoided.
4. Please use up the ECL working solution within one day after it has been configured, do not leave it until the next day, so as not to affect the accuracy of the experimental results.
5. After the use of each solution, please keep the bottle tightly capped and keep it away from light to prevent failure; especially solution B, which contains oxidizing agents and is easily reduced and become ineffective. In addition, liquid A should not be stored close to the inner wall of the refrigerator to prevent the precipitation of crystals due to the local temperature is too low. If crystals are precipitated, dissolve and mix in 37℃ warm water bath before use.
6. ECL chemiluminescence reagent kit selection refer to Table 1, primary antibody and secondary antibody recommended dilution ratio test data are from self-research antibody.
7. Refer to Table 2 for common problems and solutions for ECL chemiluminescent assays.
8. ECL chemiluminescence kit A/B solution are harmful to human body, must be careful when handling, pay attention to effective protection, avoid direct contact with the human body or inhalation of the respiratory tract.
9. For your safety and health, please wear safety glasses, gloves, or protective clothing.
Table 1: ECL Chemiluminescent Kit Selection Reference Table
| Product Name: Cat. No. | Normal ECL Chemiluminescence Kit: G2014 |
| Sensitive ECL Chemiluminescent Kit: G2074 | Hypersensitive ECL Chemiluminescence Kit: G2020 |
| Product Name: Detection limit | Normal ECL Chemiluminescence Kit: Nanogram |
| Sensitive ECL Chemiluminescent Kit: Picogram | Hypersensitive ECL Chemiluminescence Kit: Femtogram |
| Product Name: Recommended dilution ratio of primary antibody （1 mg/mL storage solution） | Normal ECL Chemiluminescence Kit: 1:1000-1:5000 |
| Sensitive ECL Chemiluminescent Kit: 1:4000-1:10000 | Hypersensitive ECL Chemiluminescence Kit: 1:10000-30000 |
| Product Name: Recommended dilution ratio of secondary antibody（1 mg/mL storage solution） | Normal ECL Chemiluminescence Kit: 1:1000-1:6000 |
| Sensitive ECL Chemiluminescent Kit: 1:3000-1:60000 | Hypersensitive ECL Chemiluminescence Kit: 1:6000-150000 |
| Product Name: Characteristics and applicability | Normal ECL Chemiluminescence Kit: Wide range, broad applicability |
| Sensitive ECL Chemiluminescent Kit: Wide range, broad applicability, high sensitivity, medium-abundance proteins, antibody-sparing | Hypersensitive ECL Chemiluminescence Kit: Very high sensitivity, low abundance proteins, low or precious antibody potency |
Table2:Common problems and Solutions of ECL Chemiluminescence Detection
Switching to a more sensitive ECL chemiluminescence kit
| C ommon Pr oblems: High background (high background or no specific bands) | Possible C auses: Primary and secondary antibody dilution without using the correct buffer and concentration |
| Solutions: Appropriately reduce the dilution ratio, reduce the concentration of primary and secondary antibodies | Blocking time or washing time is too short, or blocking solution is incorrect |
| Phosphorylated protein assays should always be closed with BSA or protein-free closures; the smaller the membrane pore size, the longer the closure and elution time should be | Inadequate incubation of primary antibody |
| Appropriately prolonged incubation time (overnight incubation at 4°C) | C ommon Pr oblems: Weak or No Signal |
| Possible C auses: Low concentration of primary and secondary antibodies used | Solutions: Increase the concentration of primary and secondary antibodies used |
| Low protein abundance | Increase sample volume |
| C ommon Pr oblems: Rapid fluorescence burst, hollow bands (ghost bands) appear | Possible C auses: The fluorescence of the target band is too strong, which rapidly consumes the luminescent substrate, and after the substrate is consumed, it will be anti-white. |
| Solutions: Reduce the amount of protein uploaded or the amount of primary and secondary antibodies | C ommon Pr oblems: Brown or yellow bands appear on the membrane |
| Possible C auses: Over-abundance of HRP in the target region generates large amounts of free radicals, resulting in oxidative inactivation of HRP | Solutions: Reduce the amount of protein uploaded or the amount of primary and secondary antibodies |
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