VCP ELISA Kit

Catalog No. ABIN7012864

Product Details VCP ELISA Kit

Target: VCP (Valosin Containing Protein (VCP))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.312 ng/mL - 20 ng/mL

Minimum Detection Limit: 0.312 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: Cell Culture Supernatant, Cell Lysate, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Valosin Containing Protein.

Sensitivity: 0.127 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 0.5 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 20ng/mL. Prepare 7 tubes containing 0.25 mL Standard Diluent and produce a double dilution series by transferring 250 µL each. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 20ng/mL, 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.312ng/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 stockDetection 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 VCP

Target: VCP (Valosin Containing Protein (VCP))

Alternative Name: Valosin Containing Protein (VCP Products)

Synonyms: ALS14 ELISA Kit, IBMPFD ELISA Kit, TERA ELISA Kit, p97 ELISA Kit, CDC48 ELISA Kit, wu:fd16d05 ELISA Kit, wu:fj63d11 ELISA Kit, p93 ELISA Kit, 3110001E05 ELISA Kit, p97/VCP ELISA Kit, 22.26 ELISA Kit, 7.12 ELISA Kit, Apple ELISA Kit, BcDNA.GM02885 ELISA Kit, BcDNA:GM02885 ELISA Kit, CG2331 ELISA Kit, DmTER94 ELISA Kit, Dmel\CG2331 ELISA Kit, I ELISA Kit, Ter94 ELISA Kit, Tera ELISA Kit, VCP ELISA Kit, VCP ATPase ELISA Kit, VCP/p97 ELISA Kit, Vcp ELISA Kit, anon-WO2004063362.65 ELISA Kit, anon-WO2004063362.67 ELISA Kit, cdc48 ELISA Kit, dTERA ELISA Kit, dVCP ELISA Kit, dmTERA ELISA Kit, dvcp ELISA Kit, l(2)03775 ELISA Kit, l(2)46CFf ELISA Kit, l(2)46CFs ELISA Kit, l(2)46Ch ELISA Kit, pTER94 ELISA Kit, ter94 ELISA Kit, vcp ELISA Kit, FANCG ELISA Kit, valosin containing protein ELISA Kit, valosin containing protein S homeolog ELISA Kit, valosin-containing protein ELISA Kit, CG2331 gene product from transcript CG2331-RA ELISA Kit, zgc:136908 ELISA Kit, transitional endoplasmic reticulum ATPase ELISA Kit, VCP ELISA Kit, vcp.S ELISA Kit, vcp ELISA Kit, Vcp ELISA Kit, Bm1_56585 ELISA Kit, LOC100136246 ELISA Kit, TER94 ELISA Kit, zgc:136908 ELISA Kit, LOC101109680 ELISA Kit

Background: IBMPFD, p97, TERA, Transitional Endoplasmic Reticulum ATPase, 15S Mg(2+)-ATPase p97 subunit

Pathways: ER-Nucleus Signaling, Positive Regulation of Endopeptidase Activity, Ubiquitin Proteasome Pathway