TUBB Kit ELISA

N° du produit ABIN6970926

Détail du produit TUBB Kit ELISA

Antigène: TUBB (Tubulin, beta (TUBB))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 0.156 ng/mL - 10 ng/mL

Seuil minimal de détection: 0.156 ng/mL

Application: ELISA

Fonction: For quantitative detection of TUBB in serum, plasma, tissue homogenates.

Type d'échantillon: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

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

Sensibilité: 0.094 ng/mL

Ingrédients:

• Pre-coated, ready to use 96-well strip plate
• Plate sealer for 96 wells
• Standard
• Sample/Standard Dilution Buffer
• Biotin-labeled Antibody (Concentrated)
• Antibody Dilution Buffer
• HRP-Streptavidin Conjugate (SABC)
• SABC Dilution Buffer
• TMB Substrate
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Matériel non inclus:

1. Microplate reader (wavelength:450nm)
2. 37 °C incubator
3. Automated plate washer
4. Precision single and multi-channel pipette and disposable tips
5. Clean tubes and Eppendorf tubes
6. Deionized or distilled water

Information d'application

Volume d'échantillon: 100 μL

Plaque: Pre-coated

Protocole:

1. Wash plate 2 times before adding Standard, Sample (diluted at least 1/2 with Sample Dilution Buffer) and Control (blank) wells!
2. Add 100 µL standard or sample to each well and incubate for 90 minutes at 37 °C.
3. Aspirate and wash plates 2 times.
4. Add 100 µL Biotin-labeled antibody working solution to each well and incubate for 60 minutes at 37 °C.
5. Aspirate and wash plates 3 times.
6. Add 100 µL SABC Working Solution into each well and incubate for 30 minutes at 37 °C.
7. Aspirate and wash plates 5 times.
8. Add 90 µL TMB Substrate Solution. Incubate 10-20 minutes at 37 °C.
9. Add 50 µL Stop Solution. Read at 450nm immediately and calculation.

Préparation des réactifs:

• Bring all reagents and samples to room temperature for 20 minutes before use.
• Wash Buffer: If crystals have formed in the concentrate, you can warm it with 40 °C water bath Concentrated Wash Buffer into 750 mL Wash Buffer with deionized or distilled water. Put unused solution back at 2-8 °C.
• Standards:
  1. Add 1 mL Sample Dilution Buffer into one Standard tube (labeled as zero tube), keep the tube at room temperature for 10 minutes and mix them thoroughly. Note: If the standard tube concentration higher than the range of the kit,please dilute it and labeled as zero tube.
  2. Label 7 EP tubes with 1/2, 1/4, 1/8, 1/16, 1/32, 1/64 and blank respectively. Add 0.3 mLof the Sample Dilution Buffer into each tube. Add 0.3 mLof the above Standard solution (from zero tube) into 1st tube and mix them thoroughly. Transfer 0.3 mL from 1st tube to 2nd tube and mix them thoroughly. Transfer 0.3 mL from 2nd tube to 3rd tube and mix them thoroughly, and so on. Sample Dilution Buffer was used for the blank control. Note: It is best to use Standard Solutions within 2 hours.
• Preparation of Biotin-labeled Antibody Working Solution:
  Prepare it within 1 hour before experiment.
  1. Calculate required total volume of the working solution: 0.1ml/well x quantity of wells. (Allow 0.1-0.2 mLmore than the total volume.)
  2. Dilute the Biotin-detection antibody with Antibody Dilution Buffer at 1:100 and mix them thoroughly. (i.e. Add 1 µL Biotin-labeled antibody into 99 µL Antibody Dilution Buffer.)
• Preparation of HRP-Streptavidin Conjugate (SABC) Working Solution:
  Prepare it within 30 minutes before experiment.
  1. Calculate required total volume of the working solution: 0.1ml/well x quantity of wells. (Allow 0.1-0.2 mLmore than the total volume.)
  2. Dilute the SABC with SABC Dilution Buffer at 1:100 and mix them thoroughly. (i.e. Add 1 µL of SABC into 99 µL of SABC Dilution Buffer.)

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).

Note:

The user should estimate the concentration of target protein in the test sample, and select a proper dilution factor to make the diluted target protein concentration fall in the optimal detection range of the kit. Dilute the sample with the provided dilution buffer, and several trials may be necessary. The test sample must be well mixed with the dilution buffer. And also standard curves and sample should be making in pre-experiment. If samples with very high concentrations, dilute samples with PBS first and then dilute the samples with Sample Dilution. The matrix components in the sample will affect the test results, which it need to be diluted at least 1/2 with Sample Dilution Buffer before testing!

Procédure de l'essai:

Washing

Manual: Discard the solution in the plate without touching the side walls. Clap the plate on absorbent filter papers or other absorbent material. Fill each well completely with 350 µL wash buffer and soak for 1 to 2 minutes, then aspirate contents from the plate, and clap the plate on absorbent filter papers or other absorbent material.
Automatic: Aspirate all wells, and then wash plate with 350 µL wash buffer. After the final wash, invert plate, and clap the plate on absorbent filter papers or other absorbent material. It is recommended that the washer shall be set for soaking 1 minute. (Note: set the height of the needles, be sure the fluid can be sipped up completely)

When diluting samples and reagents, they must be mixed completely and evenly. Before adding TMB into wells, equilibrate TMB Substrate for 30 minutes at 37 °C. It is recommended to plot a standard curve for each test.

1. Set standard, test samples (diluted at least 1/2 with Sample Dilution Buffer), control (blank) wells on the pre-coated plate respectively, and then, records their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (blank) wells!
2. Prepare Standards: Aliquot 100 µL of zero tube, 1sttube, 2ndtube, 3rdtube, 4thtube, 5thtube, 6thtube and Sample Dilution Buffer (blank) into the standard wells.
3. Add Samples: Add 100 µL of properly diluted sample into test sample wells.
4. Incubate: Seal the plate with a cover and incubate at 37 °C for 90 minutes.
5. Wash: Remove the cover and discard the plate content, and wash plate 2 times with Wash Buffer. Do NOT let the wells dry completely at any time.
6. Biotin-labeled Antibody: Add 100 µL Biotin-labeled antibody working solution into above wells (standard, test sample and blank wells). Add the solution at the bottom of each well without touching the sidewall, cover the plate and incubate at 37 °C for 60 minutes.
7. Wash: Remove the cover, and wash plate 3 times with Wash Buffer, and let the Wash Buffer stay in the wells for 1-2 minutes each time.
8. HRP-Streptavidin Conjugate (SABC): Add 100 µL of SABC Working Solution into each well, cover the plate and incubate at 37 °C for 30 minutes.
9. Wash: Remove the cover and wash plate 5 times with Wash Buffer, and let the wash buffer stay in the wells for 1-2 minutes each time.
10. TMB Substrate: Add 90 µL TMB Substrate into each well, cover the plate and incubate at 37 °C in dark within 10-20 minutes. (Note: The reaction time can be shortened or extended according to the actual color change, but not more than 30 minutes. You can terminate the reaction when apparent gradient appeared in standard wells.)
11. Stop: Add 50 µL Stop Solution into each well. The color will turn yellow immediately. The adding order of Stop Solution should be as the same as the TMB Substrate Solution.
12. OD Measurement: Read the O.D. absorbance at 450nm in Microplate Reader immediately after adding the stop solution.

Regarding calculation, (the relative O.D.450) = (the O.D.450 of each well) - (the O.D.450 of blank well). The standard curve can be plotted as the relative O.D.450 of each standard solution (Y) vs. the respective concentration of the standard solution (X). The target concentration of the samples can be interpolated from the standard curve. It is recommended to use some professional software to do this calculation, such as Curve Expert 1.3 or 1.4.

Note: If the samples measured were diluted, multiply the dilution factor to the concentrations from interpolation to obtain the concentration before dilution.

Précision du teste: Intra-Assay: CV<8%
Inter-Assay: CV<10%

Restrictions: For Research Use only

Stockage

Stock: 4 °C,-20 °C

Stockage commentaire:

1. For unopened kit: All the reagents should be kept according to the labels on vials. The Reference Standard and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
2. For used kit: When the kit is used, the remaining reagents 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 zip-seal the foil pouch.

Date de péremption: 6 months

Détail du antigène

Antigène: TUBB (Tubulin, beta (TUBB))

Autre désignation: Tubulin beta chain (TUBB Produits)

Synonymes: M40 Kit ELISA, OK/SW-cl.56 Kit ELISA, TUBB1 Kit ELISA, TUBB5 Kit ELISA, XLOT Kit ELISA, m40 Kit ELISA, ok/sw-cl.56 Kit ELISA, tubb1 Kit ELISA, tubb5 Kit ELISA, TUBB2A Kit ELISA, TUBB2B Kit ELISA, TUBB Kit ELISA, 1t Kit ELISA, B1t Kit ELISA, BETA 56D Kit ELISA, CG9277 Kit ELISA, DTB2 Kit ELISA, Dmbeta1 Kit ELISA, Dmel\\CG9277 Kit ELISA, T Kit ELISA, Tub Kit ELISA, Tubulin Kit ELISA, beta Kit ELISA, beta-Tub Kit ELISA, beta-Tub56D Kit ELISA, beta-tub Kit ELISA, beta-tubulin56D Kit ELISA, beta1 Kit ELISA, beta1-Tubulin Kit ELISA, beta1-tub Kit ELISA, beta1Tub Kit ELISA, beta1t Kit ELISA, beta1tub Kit ELISA, beta56D Kit ELISA, betaTub Kit ELISA, betaTub1 Kit ELISA, beta[[1]] tubulin Kit ELISA, beta[[1]]-tubulin Kit ELISA, betatub(56D) Kit ELISA, 143391_i_at Kit ELISA, 3t Kit ELISA, B3t Kit ELISA, BETA 60D Kit ELISA, CG3401 Kit ELISA, D.m.BETA-60D Kit ELISA, DTB3 Kit ELISA, Dmbeta3 Kit ELISA, Dmel\\CG3401 Kit ELISA, Tub60D Kit ELISA, beta-Tub60D Kit ELISA, beta-Tub6D Kit ELISA, beta3 Kit ELISA, beta3 TU Kit ELISA, beta3-Tub Kit ELISA, beta3-tubulin Kit ELISA, beta3TUB Kit ELISA, beta3Tub Kit ELISA, beta3t Kit ELISA, beta60C Kit ELISA, betaTub3 Kit ELISA, betaTub60C Kit ELISA, beta[[3]] tubulin Kit ELISA, beta[[3]]-Tub Kit ELISA, beta[[3]]-tubulin Kit ELISA, betatub60D Kit ELISA, p50 Kit ELISA, p50/tubulin Kit ELISA, p53 Kit ELISA, p53/tubulin Kit ELISA, 2t Kit ELISA, B2t Kit ELISA, BETA 85D Kit ELISA, BETA2 Kit ELISA, CG9359 Kit ELISA, D.m.BETA-85D Kit ELISA, DTB4 Kit ELISA, Dmbeta2 Kit ELISA, Dmel\\CG9359 Kit ELISA, beta(2)Tu Kit ELISA, beta(2)Tub Kit ELISA, beta-Tub85D Kit ELISA, beta-tub85D Kit ELISA, beta2 Kit ELISA, beta2-tub Kit ELISA, beta2t Kit ELISA, beta2tub Kit ELISA, beta85D Kit ELISA, betaTub2 Kit ELISA, beta[[2]]-tubulin Kit ELISA, ms(3)KK[D] Kit ELISA, 4t Kit ELISA, B4t Kit ELISA, BETA 98B Kit ELISA, CG4869 Kit ELISA, DTB1 Kit ELISA, Dmbeta4 Kit ELISA, Dmel\\CG4869 Kit ELISA, beta-Tub97EF Kit ELISA, beta4 Kit ELISA, beta4t Kit ELISA, beta97F Kit ELISA, betaTub4 Kit ELISA, betaTub98BC Kit ELISA, betaTub98C Kit ELISA, Tub1 Kit ELISA, bmtub1 Kit ELISA, Tub3 Kit ELISA, bmtub3 Kit ELISA, Tub2 Kit ELISA, bmtub2 Kit ELISA, Tub4 Kit ELISA, bmtub4 Kit ELISA, LOC100101153 Kit ELISA, tubulin beta class I Kit ELISA, putative cobalt-zinc-cadmium outer membrane resistance protein Kit ELISA, Tubulin beta chain Kit ELISA, tubulin beta-2A chain Kit ELISA, beta-Tubulin at 56D Kit ELISA, beta-Tubulin at 60D Kit ELISA, beta-Tubulin at 85D Kit ELISA, beta-Tubulin at 97EF Kit ELISA, beta-tubulin Kit ELISA, tubulin beta 6 class V Kit ELISA, beta tubulin Kit ELISA, tubulin beta class I L homeolog Kit ELISA, tubulin, beta 5 class I Kit ELISA, TUBB Kit ELISA, czcC Kit ELISA, tbb-6 Kit ELISA, tubb Kit ELISA, LOC462399 Kit ELISA, betaTub56D Kit ELISA, betaTub60D Kit ELISA, betaTub85D Kit ELISA, betaTub97EF Kit ELISA, Tub1 Kit ELISA, Tub3 Kit ELISA, Tub2 Kit ELISA, Tub4 Kit ELISA, TVAG_525430 Kit ELISA, TVAG_062880 Kit ELISA, TVAG_456920 Kit ELISA, LOC100101153 Kit ELISA, TUBB6 Kit ELISA, Tb927.1.2330 Kit ELISA, Tb927.1.2350 Kit ELISA, Tb927.1.2370 Kit ELISA, Tb927.1.2390 Kit ELISA, tubb.L Kit ELISA, Tubb Kit ELISA, LOC693049 Kit ELISA, LOC443015 Kit ELISA, Tubb5 Kit ELISA

Sujet: TUBβ, β-Tubulin, TUBB, OK, SW-cl.56, TUBB5, Tubulin beta chain, Tubulin beta-5 chain

UniProt: P07437

Pathways: Dynamique des Microtubules, M Phase