beta Actin Kit ELISA (Actin, beta)

Details for Product ACTB ELISA Kit No. ABIN6574121, Fournisseur: Connectez-vous pour afficher
Antigène
  • BRWS1
  • PS1TP5BP1
  • ACTA1
  • Actx
  • E430023M04Rik
  • beta-actin
  • ps1tp5bp1
  • act-2
  • actb
  • B-actin
  • LOC100174883
  • LOC100304816
  • A
  • ACT5C
  • Ac5C
  • Act
  • Act-5C
  • Act5
  • Act5c
  • Actin
  • Actin/BAP47
  • Actin5C
  • BAP47
  • Bap47
  • CG4027
  • Dmel\\CG4027
  • M32055
  • T11
  • act
  • act 5C
  • act5C
  • actin
  • actin5C
  • anon-EST:fe2D2
  • beta-actin/Bap47
  • cyt5C
  • l(1)G0009
  • l(1)G0010
  • l(1)G0025
  • l(1)G0079
  • l(1)G0117
  • l(1)G0177
  • l(1)G0245
  • l(1)G0330
  • l(1)G0420
  • l(1)G0486
  • Bact
  • ACT-5
  • ACTB
  • B-ACTZF
  • actba
  • bact
  • bactin1
  • bactzf
  • wu:fd18f05
  • Beta-actin
  • actin beta
  • actin, beta
  • cytoplasmic actin OlCA1
  • beta-actin
  • Actin 5C
  • actin gamma 1
  • actin, beta 1
  • actin, beta L homeolog
  • beta actin
  • actin, cytoplasmic 1
  • ACTB
  • Actb
  • actb
  • LOC100174883
  • LOC100304816
  • Act5C
  • ACTG1
  • actb1
  • actb.L
  • LOC100136352
  • LOC109073280
Reactivité
Humain, Souris, Rat (Rattus)
Alternatives
Kits with alternative reactivity to:
19
19
14
4
4
3
2
2
2
2
2
1
1
Méthode de détection
Chemiluminescent
Type de méthode
Sandwich ELISA
Gamme de detection
0.14 ng/mL - 100 ng/mL
Seuil minimal de détection
0.14 ng/mL
Application
ELISA
Options
Fournisseur
Connectez-vous pour afficher
N° du produit (Fournisseur)
Connectez-vous pour afficher
Fonction The Chemiluminescent Immunoassay kit is designed for the in vitro sensitive quantitative measurement of ACTb in tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Type d'échantillon Cell Culture Supernatant, Cell Lysate, Tissue Homogenate
Analytical Method Quantitative
Méthode de détection Chemiluminescent
Specificité

This assay has high sensitivity and excellent specificity for detection of Actin Beta (ACTb).
No significant cross-reactivity or interference between Actin Beta (ACTb) and analogues was observed.

Sensibilité 0.05 ng/mL
Ingrédients
  • Pre-coated, ready to use 96-well strip plate
  • Plate sealer for 96 wells
  • Standard
  • Standard Diluent
  • Assay Diluent A
  • Assay Diluent B
  • Standard
  • Detection Reagent A
  • Detection Reagent B
  • Substrate A
  • Substrate B
  • Wash Buffer (30 x concentrate)
  • Instruction manual
Matériel non inclus
  • Luminometer capable of reading 96-well microplates with the following parameters: lag time 30.0s, read time 1.0 s/well .
  • Precision single or multi-channel pipettes and pipette tips with disposable tips.
  • Eppendorf Tubes for diluting samples.
  • Deionized or distilled water.
  • Absorbent paper for blotting the microtiter plate.
  • Container for Wash Solution
Plasmids, Primers & others Plasmids, Primers & others beta Actin products on genomics-online (e.g. as negative or positive controls)
Antigène
Autre désignation Actin Beta (ACTB ELISA Kit Extrait)
Pathways Myometrial Relaxation and Contraction, Cell-Cell Junction Organization, Maintenance of Protein Location, Phototransduction
Indications d'application
  • Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
  • The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
  • Kits from different batches may be a little different in detection range, sensitivity and color developing time.
  • Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
  • Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
  • There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
  • Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
  • Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
  • Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
  • Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.
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 The microplate provided in this kit has been pre-coated with an antibody specific to Actin Beta (ACTb). Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to Actin Beta (ACTb). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Actin Beta (ACTb) level in the sample or standard.,
Préparation de l'échantillon
  • We are only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.
  • Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments. Sample should be diluted by 0.01Mol/L PBS(PH=7.0-7.2).
  • If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
  • Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts from certain chemicals.
  • Due to the possibility of mismatching between antigen from other origin and antibody used in our kits (e.g.antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
  • Influenced by the factors including cell viability, cell number or sampling time, samples from cell culture supernatant may not be detected by the kit.
  • Fresh samples without long time storage is recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.
Procédure de l'essai
  • 1. Determine wells for diluted standard, blank and sample. Prepare 7 wells for standard, 1 well for blank. Add 100 µL each of dilutions of standard (read Reagent Preparation), blank and samples into the appropriate wells. Cover with the Plate sealer. Incubate for 2 hours at 37 °C.
  • 2. Remove the liquid of each well, don't wash.
  • 3. Add 100 µL of Detection Reagent A working solution to each well. Incubate for 1 hour at 37 °C after covering it with the Plate sealer.
  • 4. Aspirate the solution and wash with 350 µL of 1× Wash Solution to each well using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher, and let it sit for 1~2 minutes. Remove the remaining liquid from all wells completely by snapping the plate onto absorbent paper. Repeat 3 times. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against absorbent paper.
  • 5. Add 100 µL of Detection Reagent B working solution to each well. Incubate for 30 minutes at 37 °C after covering it with the Plate sealer.
  • 6. Repeat the aspiration/wash process for five times as conducted in step 4.
  • 7. Add 100 µL of Substrate working Solution to each well. Cover with a new Plate sealer. Incubate for 5-10 minutes at 37 °C (Don't exceed 10 minutes). Protect from light.
  • 8. Measure the chemiluminescence signal in a microplate luminometer or as appropriate for the instrument used.
Note:
  • 1. Assay preparation: Keep appropriate numbers of wells for 1 experiment and remove extra wells from microplate. Rest wells should be resealed and stored at -20 °C.
  • 2. Samples or reagents addition: Please use the freshly prepared Standard. Please carefully add samples to wells and mix gently to avoid foaming. Do not touch the well wall. For each step in the procedure, total dispensing time for addition of reagents or samples to the assay plate should not exceed 10 minutes. This will ensure equal elapsed time for each pipetting step, without interruption. To avoid cross-contamination, change pipette tips between additions of standards, samples, and reagents. Also, use separated reservoirs for each reagent.
  • 3. Incubation: To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. Once reagents are added to the well strips, DO NOT let the strips DRY at any time during the assay. Incubation time and temperature must be controlled.
  • 4. Washing: The wash procedure is critical. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting and remove any drop of water and fingerprint on the bottom of the plate. Insufficient washing will result in poor precision and false elevated absorbance reading.
  • 5. For Substrate A and B, please protect it from light.
  • 6. Relative light units (RLUs) may differ from different luminometers.The Immunoassay was optimized using a Beijing Hamamatsu luminometer. Other instruments may require settings to be adjusted.
  • 7. Relative light units may vary within the 10 minute reading window.
Précision du teste

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Actin Beta (ACTb) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Actin Beta (ACTb) 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
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.
Conseil sur la manipulation The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Stock 4 °C
Stockage commentaire
  • For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
  • For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
    Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
  • For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.
Date de péremption 6 months
Images (Fournisseur)
ELISA image for Actin, beta (ACTB) ELISA Kit (ABIN6574121) Typical standard curve
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