PPIA ELISA Kit

Catalog No. ABIN6974526

Product Details PPIA ELISA Kit

Target: PPIA (Peptidylprolyl Isomerase A (Cyclophilin A) (PPIA))

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 125 pg/mL - 8000 pg/mL

Minimum Detection Limit: 125 pg/mL

Application: ELISA

Purpose: For the quantitative determination of rat cyclophilin A (CyPA) concentrations in serum, plasma, tissue homogenates, cell lysates.

Sample Type: Cell Lysate, Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of rat CyPA. No significant cross-reactivity or interference between rat CyPA and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between rat CyPA and all the analogues, therefore, cross reaction may still exist.

Sensitivity: 31.25 pg/mL

Components:

• Assay plate
• Standard
• HRP-avidin (100 x concentrate)
• Biotin-antibody (100 x concentrate)
• Sample Diluent
• HRP-avidin Diluent
• Biotin-antibody Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 100 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Reagent Preparation:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. Wash Buffer(1x)- If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 8000 pg/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (8000 pg/mL). Sample Diluent serves as the zero standard (0 pg/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. 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): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date. May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Expiry Date: 6 months

References for PPIA ELISA Kit (ABIN6974526)

Tang, Wang, Ma, Guan, Pan, Yao, Chen, Li, Yang, Zhu, Tu: "Simvastatin attenuated rat thoracic aorta remodeling by decreasing ROCK2‑mediated CyPA secretion and CD147‑ERK1/2‑cyclin pathway." in: Molecular medicine reports, Vol. 16, Issue 6, pp. 8123-8129, (2018) (PubMed).

Target Details for PPIA

Target: PPIA (Peptidylprolyl Isomerase A (Cyclophilin A) (PPIA))

Alternative Name: peptidylprolyl isomerase A (cyclophilin A) (PPIA Products)

Synonyms: Cyp1 ELISA Kit, PPIA-2 ELISA Kit, fa93g09 ELISA Kit, ppia ELISA Kit, wu:fa93g09 ELISA Kit, wu:fb05e11 ELISA Kit, wu:fb13h02 ELISA Kit, wu:fb15a05 ELISA Kit, CYPA ELISA Kit, CypA ELISA Kit, Cypa ELISA Kit, CYPH ELISA Kit, CYCA ELISA Kit, CyP-A ELISA Kit, MGC75715 ELISA Kit, 2700098C05 ELISA Kit, Cphn ELISA Kit, CyP-18 ELISA Kit, MT-ND1 ELISA Kit, MTND1 ELISA Kit, NADH1 ELISA Kit, ND1 ELISA Kit, cypA ELISA Kit, PPIase A ELISA Kit, ppial ELISA Kit, wu:fj18g05 ELISA Kit, zgc:73102 ELISA Kit, zgc:86688 ELISA Kit, peptidylprolyl isomerase A ELISA Kit, peptidylprolyl isomerase Aa (cyclophilin A) ELISA Kit, peptidylprolyl isomerase A (cyclophilin A) L homeolog ELISA Kit, cyclophilin A ELISA Kit, cyclophilin a ELISA Kit, peptidylprolyl isomerase A (cyclophilin A) ELISA Kit, peptidylprolyl isomerase Ab (cyclophilin A) ELISA Kit, PPIA ELISA Kit, ppiaa ELISA Kit, ppia.L ELISA Kit, Cypa ELISA Kit, CNB01230 ELISA Kit, CNB01290 ELISA Kit, Tc00.1047053506925.300 ELISA Kit, Tb11.03.0250 ELISA Kit, CYPA ELISA Kit, Ppia ELISA Kit, ppia ELISA Kit, ppiab ELISA Kit

Background:

Abbreviation: PPIA

Alias: CYPA, CYPH, MGC117158, MGC12404, MGC23397, PPIase A|T cell cyclophilin|cyclosporin A-binding protein|peptidyl-prolyl cis-trans isomerase A|peptidylprolyl isomerase A|rotamase A

UniProt: P10111