Overview

  • Product name

    Mouse ICAM1 ELISA Kit (CD54)
    See all ICAM1 kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 20 pg/ml
  • Range

    24.69 pg/ml - 6000 pg/ml
  • Recovery

    97 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 93.54 84% - 103%
    Serum 94.56 84% - 103%
    Plasma 115.2 97% - 127%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Abcam’s ICAM1 Mouse ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Mouse ICAM1 in serum, plasma and cell culture supernatants.

    This assay employs an antibody specific for mouse ICAM1 coated on a 96- well plate. Standards and samples are pipetted into the wells and ICAM1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse ICAM1 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of ICAM1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    400X HRP-Streptavidin Concentrate 1 x 200µl
    5X Assay Diluent B 1 x 15ml
    5X Assay Diluent D 1 x 15ml
    Biotinylated anti-Mouse ICAM1 2 vials
    ICAM1 Microplate (12 x 8 wells) 1 unit
    Recombinant Mouse ICAM1 Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

  • Relevance

    ICAM1 is a 85-110 kDa single chain type 1 integral membrane glycoprotein with an extracellular domain of five immunoglobulin superfamily repeats, a transmembrane region and a cytoplasmic domain. It shares considerable amino acid sequence homology with ICAM3 and with ICAM2. ICAM1 is expressed by activated endothelial cells. It is detected on cells of many other lineages (e.g. epithelial cells, fibroblasts, chondrocytes, B lymphocytes, T lymphocytes (low), monocytes, macrophages, dendritic cells and neutrophils), with lower levels that increase in inflammation. ICAM1 is also detected in some carcinoma and melanoma cells. Soluble ICAM1 is detectable in the plasma and is elevated in patients with various inflammatory syndromes. It is the receptor for rhinoviruses and malaria.
  • Cellular localization

    Membrane
  • Alternative names

    • BB2
    • CD 54
    • CD54
    • CD54 antigen
    • Cell surface glycoprotein P3.58
    • Human rhinovirus receptor
    • ICAM 1
    • ICAM-1
    • Intercellular adhesion molecule 1
    • Major group rhinovirus receptor
    • MALA2
    • MGC6195
    • MyD10
    • P3.58
    • sICAM-1
    • Surface antigen of activated B cells
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab100688 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Representative Standard Curve

Protocols

References

This product has been referenced in:

  • Wang Y  et al. Role of the Rho/ROCK signaling pathway in the protective effects of fasudil against acute lung injury in septic rats. Mol Med Rep 18:4486-4498 (2018). Read more (PubMed: 30221694) »
  • Jing SH  et al. Raf Kinase Inhibitor Protein (RKIP) Inhibits Tumor Necrosis Factor-a (TNF-a) Induced Adhesion Molecules Expression in Vascular Smooth Muscle Bells by Suppressing (Nuclear Transcription Factor-?B (NF-kappaB) Pathway. Med Sci Monit 23:4789-4797 (2017). ELISA ; Mouse . Read more (PubMed: 28983072) »
See all 4 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
Answer

Thank you for contacting us. Althought this particular kit has not, to our knowledge, been tested with tissue homogenates, it should be compatible.

We recommend diluting the samples at least 5-fold with Assay Diluent D to minimize any effects of the detergents in the lysis buffer. The samples may need to be diluted further but this would need to be determined by the experimenter empirically.

In brief, a lysis buffer must meet the following specifications:

A) has relatively low salt content (700 mM or less)

B) does not contain sodium azide

C) does not contain >0.1% SDS

D) does not contain >10 mM reducing agents (beta-mercaptoethanol or dithiothreitol)

For the original lysate, in general you want to obtain a stock concentration at least 1 mg/ml protein, preferably more, to achieve 50-500 ug/ml after dilution. This range should just be used as a starting point and the optimal concentration would need to be determined empirically.

Here are more general instructions from the laboratory that developed this kit.

1) Avoid using 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 reducing agents, such as dithiothreitol or mercaptoethanols.

In general, we strongly recommend that you add some type of protease inhibitor “cocktail” to the lysis buffer prior to homogenization. Since susceptibility to proteolytic cleavage and the type of proteases present in the lysate vary, we do not recommend a specific product. Instead, your choice of which combination of protease inhibitors to use should be based upon a literature search for your protein(s) of interest and/or tissue or cell type. Phosphatase inhibitors may be used, but are not necessary unless the antibodies used in the kit specifically recognize phosphorylated (activated) forms of the protein.

Choices of the method for lysis and homogenization include glass-bead “smash,” douncing, freeze-thaw, sonication and crushing frozen tissue with a mortar and pestle, or even a combination of these. There is no one “best method” for all sample types, but some are better than others for some 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, spin down the lysates to remove cell/tissue debris (5 min @ 10000 x g or 10 min @ 5000 x g) and save the supernatant. Lysates should be frozen as soon as possible, and stored at -20°C (or -80°C, if possible). Spin them again before incubating with the antibody array. Determine the protein concentration of your lysates (for example the bicinchoninic acid (BCA) assay) and normalize the volume of each sample used to deliver the same amount of total protein for each assay.

Since different cells and tissues may contain different amounts of protein, as starting point, we suggest using 500 uL of lysis buffer per 1e6 cells or 10 mg tissue. You may have to adjust this based upon your results. Your target for total protein concentration of the homogenate should be at least 1000 ug/mL, but 2000 ug/mL would be better.

Read More

Answer

Thank you for contacting us.

The following ELISA kits for mouse ICAM, VCAM, IL-6 and TNF-alpha are suitable for mouse samples:

ab100688 (https://www.abcam.com/ICAM1-Mouse-ELISA-Kit-1-x-96-Well-Plate-ab100688.html)
ab100713 (https://www.abcam.com/IL6-Mouse-ELISA-Kit-1-x-96-Well-Plate-ab100713.html) was tested on tissue as well.
ab100747 (https://www.abcam.com/TNF-alpha-Mouse-ELISA-Kit-1-x-96-Well-Plate-ab100747.html)
ab100750 (https://www.abcam.com/VCAM1-Mouse-ELISA-Kit-1-x-96-Well-Plate-ab100750.html)

Please let me know if these are kits you were refering to.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

Thank you for your call earlier this week and for your patience while I have looked into your enquiry. I contacted the lab to ask about cross-reactivity of our mouse ICAM1 and VCAM ELISA kits with the other CAM's, and unfortunately the antibodies in these kits are not expected to cross-react with VCAM, ICAM, and PCAM. I am sorry that we do not have the product you are looking for, but if you have any questions or need anything further please let me know.

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