CDC42 Kit ELISA

N° du produit ABIN6954549

Détail du produit CDC42 Kit ELISA

Antigène: CDC42 (Cell Division Cycle 42 (GTP Binding Protein, 25kDa) (CDC42))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 0.15 ng/mL - 10 ng/mL

Seuil minimal de détection: 0.15 ng/mL

Application: ELISA

Fonction: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of CDC42 in human tissue homogenates, cell lysates.

Type d'échantillon: Cell Lysate, Tissue Homogenate

Analytical Method: Quantitative

Specificité: This assay has high sensitivity and excellent specificity for detection of Cell Division Cycle Protein 42 (CDC42)

Sensibilité: 0.055 ng/mL

Ingrédients:

• 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

Information d'application

Commentaires:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Volume d'échantillon: 100 μL

Durée du test: 3 h

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Préparation des réactifs:

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 40 ng/mL. Firstly dilute the stock solution to 10 ng/mL and the diluted standard serves as the highest standard (10 ng/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, 0.156 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/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 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 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 standards 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 one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Préparation de l'échantillon:

• 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).

Précision du teste: 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

Stockage

Précaution d'utilisation: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Stock: 4 °C/-20 °C

Stockage commentaire:

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.

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Date de péremption: 6 months

Détail du antigène

Antigène: CDC42 (Cell Division Cycle 42 (GTP Binding Protein, 25kDa) (CDC42))

Autre désignation: Cell Division Cycle Protein 42 (CDC42) (CDC42 Produits)

Synonymes: CDC42Hs Kit ELISA, G25K Kit ELISA, AI747189 Kit ELISA, AU018915 Kit ELISA, cdc42a Kit ELISA, wu:fb07d08 Kit ELISA, wu:fj62g06 Kit ELISA, zgc:63918 Kit ELISA, Xcdc42 Kit ELISA, cdc42hs Kit ELISA, g25k Kit ELISA, CDC42 Kit ELISA, CG12530 Kit ELISA, Cdc42Dm Kit ELISA, D-CDC42 Kit ELISA, D-Cdc42 Kit ELISA, D-cdc42 Kit ELISA, DCDC42 Kit ELISA, DCdc42 Kit ELISA, Dcdc42 Kit ELISA, Dm Cdc42 Kit ELISA, DmCDC42 Kit ELISA, Dmcdc42 Kit ELISA, Dmel\\CG12530 Kit ELISA, cdc42 Kit ELISA, cell division cycle 42 Kit ELISA, Cell division control protein 42 homolog Kit ELISA, cell division control protein 42 homolog Kit ELISA, cell division cycle 42 L homeolog Kit ELISA, CG12530 gene product from transcript CG12530-RC Kit ELISA, Rho family GTPase Kit ELISA, CDC42 Kit ELISA, Cdc42 Kit ELISA, cdc42 Kit ELISA, LOC100636735 Kit ELISA, LOC100637699 Kit ELISA, LOC100638834 Kit ELISA, cdc42.L Kit ELISA

Pathways: Signalisation MAPK, Dynamique des Microtubules, Signalisation RTK, Signalisation WNT, TCR Signaling, EGFR Signaling Pathway, Regulation of Actin Filament Polymerization, Regulation of Muscle Cell Differentiation, Cell-Cell Junction Organization, Maintenance of Protein Location, Skeletal Muscle Fiber Development, Signaling Events mediated by VEGFR1 and VEGFR2, EGFR Downregulation, VEGF Signaling