Human Obesity Antibody Array (40 Targets) - Quantitative (ab197417)

Overview

  • Product name

    Human Obesity Antibody Array (40 Targets) - Quantitative
    See all Obesity Antibody Array antibody arrays
  • Sample type

    Serum, Plasma, Other biological fluids, Cell Lysate, Cell culture media, Tissue Lysate
  • Assay type

    Quantitative
  • Assay time

    6h 00m
  • Species reactivity

    Reacts with: Human
  • Product overview

    ab197417 can be used for quantitative measurement of 40 Human cytokines. Suitable for serum, plasma, cell culture media, other body fluids, cell and tissue lysates.
    Targets names: Adiponectin, Adipsin, AgRP, ANGPTL4, BDNF, Chemerin, CRP, GH, IFNγ, IGFBP-1, IGFBP-2, IGF-I, IL-10, IL-12p40, IL-12p70, IL-1β, IL-1ra, IL-6, IL-8, Insulin, Leptin, Lipocalin-2, MSPα, OPG, PAI-1, PDGF-BB, Pepsinogen 1, Pepsinogen 2, Procalcitonin, Prolactin, RANTES, RBP4, Resistin, SAA, TGFβ1, TSP-1, TNF RI, TNF RII, TNFα and VEGF.
    Learn more about quantitative antibody arrays

  • Notes

    Quantitative antibody arrays can be used to quantitate up to 40 cytokines with as little as 50 µL of sample. Arrays are available for 400 human and 200 mouse proteins. 

    Each glass slide is spotted with 16 identical antibody arrays and is provided with a 16 well gasket to allow separate samples to be applied to each array. Each antibody is spotted in quadruplicate in each array. For reproducible quantitation, eight of the arrays are used with a cocktail of protein standards to produce a standard curve. The same 8 standard curve arrays can be used across multiple slides, allowing measurement of 22 experimental samples with 2 slides, 36 with 3 slides etc. 

    For high through-put, 4 slides can be nested into a tray matching a standard microplate, allowing automated processing with a liquid handling workstation. 

    Array processing can be completed within one working day (see workflow diagram below) and arrays can be analyzed with a wide number of laser glass slide array / gene microarray scanners. If you don’t have a suitable scanner then we recommend our membrane antibody arrays, which can be analyzed with any WB chemiluminescent reader. 

    Learn more about quantitative antibody arrays 

    Learn more about membrane antibody arrays

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 8 tests 22 units 50 tests
    20X Wash Buffer I 2 x 30ml 3 x 30ml 6 x 30ml
    20X Wash Buffer II 1 x 30ml 1 x 30ml 2 x 30ml
    Cy3 equivalent dye-conjugated Streptavidin 1 x 5µl 2 x 5µl 4 x 5µl
    Human Obesity Array (40 T) Biotinylated Antibody Cocktail 1 unit 2 units 4 units
    Human Obesity Array (40 T) Glass Slide 1 slide 2 slides 4 slides
    Human Obesity Array (40 T) Lyophilized Cytokine Standard Mix 1 vial 2 vials 4 vials
    Sample Diluent 1 x 15ml 1 x 15ml 2 x 15ml
    Slide Washer/Dryer (30 mL Centrifuge Tube) 1 unit 1 unit 2 units

Images

  • Assay Summary for Human Obesity Antibody Array - Quantitative (40 targets) (ab197417).

  • Array Map for Human Obesity Antibody Array - Quantitative (40 targets) (ab197417).

  • Standard Curve obtained with Abcam Human Obesity Antibody Array - Quantitative (40 targets) (ab197417).

Protocols

References

ab197417 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Answer

Your customer can homogenized the tissue of interest in their own lysis buffer, such as RIPA or other formulations optimized for immunoprecipitation. We can provide 10X RIPA Buffer (ab156034).



Please follow the guidelines on lysis buffer composition shown below:


1) Avoid using >0.1% SDS or other strongly denaturing detergents. In general, non-ionic detergents such as Triton X-100 or NP-40 are best, although zwitterionic detergents such as CHAPS, or mild ionic detergents such as sodium deoxycholate will work.


2) Use no more than 2% v/v total detergent


3) Avoid the use of sodium azide


4) Avoid using >10 mM reducing agents, such as dithiothreitol or mercaptoethanols



We strongly recommend adding a protease inhibitor cocktail to the lysis buffer prior to homogenization.


Choices of the method for lysis and homogenization include glass-bead “smash,” douncing, freezethaw, sonication and crushing frozen tissue with a mortar and pestle, or even a combination of these. There is no best method for all sample types; your choice of method should be made following a brief search of the literature to see how samples similar to yours have been prepared in previous investigations.


After homogenization, centrifuge the lysates to remove cell/tissue debris (5 min @ 10,000 x g or 10min @ 5,000 x g) and save the supernatant. Unless testing fresh, lysates should be frozen as soon as possible and stored at -20°C (or -80°C, if possible). Centrifuge them again before incubating with any immunoassay. Next, determine the protein concentration of your lysates using a total protein assay not inhibited by detergents (such as the Bicinchoninic acid (BCA) assay) and normalize the volume of each sample used to deliver the same amount of total protein for each assay.


Note: The Bradford assay is not recommended as it can be inhibited by the presence of detergents.


Since different cells and tissues may contain different amounts of protein, as starting point, we suggest using 500 μL of lysis buffer per 10 mg tissue. You may have to adjust this based upon your results. Your target total protein concentration of the homogenate should be at least 1,000 μg/mL, but 2,000 μg/mL or more would be better.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up