ATG5 ELISA Kit

Catalog No. ABIN7012903

Product Details ATG5 ELISA Kit

Target: ATG5 (ATG5 Autophagy Related 5 (ATG5))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.78 ng/mL - 50 ng/mL

Minimum Detection Limit: 0.78 ng/mL

Application: ELISA

Purpose: The kit is a small sample sandwich enzyme immunoassay for in vitro quantitative measurement in various sample types.

Sample Type: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Autophagy Related Protein 5.

Sensitivity: 0.28 ng/mL

Grade: Small Sample

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Sample Volume: 25 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Add 25μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 25μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 25μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 25μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 20μL Stop Solution. Read at 450nm immediately.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 200ng/mL. Firstly dilute the stock solution to 50ng/mL and the diluted standard serves as the highest standard (50ng/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and produce a double dilution series by transferring 500 µL each. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 50ng/mL, 25ng/mL, 12.5ng/mL, 6.25ng/mL, 3.12ng/mL, 1.56ng/mL, 0.78ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0ng/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 10 mL of Wash Solution concentrate (30x) with 290 mL of deionized or distilled water to prepare 300 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for once pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. Prepare Substrate working Solution within 15 minutes before assay.
6. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
7. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof 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 therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Restrictions: For Research Use only

Handling

Storage: 4 °C/-20 ° C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

Expiry Date: 6 months

Target Details for ATG5

Target: ATG5 (ATG5 Autophagy Related 5 (ATG5))

Alternative Name: Autophagy Related Protein 5 (ATG5 Products)

Synonyms: APG5 ELISA Kit, APG5-LIKE ELISA Kit, APG5L ELISA Kit, ASP ELISA Kit, hAPG5 ELISA Kit, ATG5 ELISA Kit, CG1643 ELISA Kit, DmAtg5 ELISA Kit, Dmel\\CG1643 ELISA Kit, atg5 ELISA Kit, 2010107M05Rik ELISA Kit, 3110067M24Rik ELISA Kit, AW319544 ELISA Kit, Apg5l ELISA Kit, Atg5l ELISA Kit, C88337 ELISA Kit, Paddy ELISA Kit, apg5l ELISA Kit, zgc:100934 ELISA Kit, ATATG5 ELISA Kit, AUTOPHAGY 5 ELISA Kit, MKP11.20 ELISA Kit, MKP11_20 ELISA Kit, autophagy related 5 ELISA Kit, Autophagy-related 5 ELISA Kit, ATG5 autophagy related 5 homolog (S. cerevisiae) ELISA Kit, autophagy related 5 L homeolog ELISA Kit, similar to S. cerevisiae ATG5 (YPL149W) which nucleates preautophagosome formation as a conjugate with Atg12 ELISA Kit, autophagy protein Apg5 family ELISA Kit, ATG5 ELISA Kit, Atg5 ELISA Kit, atg5 ELISA Kit, atg5.L ELISA Kit, APG5 ELISA Kit

Background: ASP, APG5, APG5L, hAPG5, Apoptosis-specific protein, Autophagy protein 5

Pathways: Activation of Innate immune Response, Production of Molecular Mediator of Immune Response, Autophagy