Western blot analysis was performed on whole cell extracts (30 ug lysate) of Raji (Lane 1), Mouse Spleen (Lane 2), Mouse Lung (lane 3), Rat Lung (lane 4) Mouse Kidney (lane 5) and Rat Kidney (lane 6). The blots were probed with ABfinity™ Anti-ICAM-1 Rabbit Monoclonal Antibody (Product# 701254, 1-2 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 ug/ml, 1:2500 dilution). A 57 kDa band corresponding to ICAM-1 was observed across cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 504–517 of human Intercellular adhesion molecule-1|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Western Blot (WB)||1-2 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
This antibody is predicted to react with mouse, rat, non-human primate and rabbit based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Intercellular adhesion molecule-1 is a member of the immunoglobulin superfamily, and is critical for the adhesion and transmigration of leukocytes from blood vessels and into tissues. ICAM-1 is constitutively expressed on endothelial cells, but levels increase in response to proinflammatory cytokines. It is also expressed on smooth muscle cells, resulting in leukocyte accumulation, and activation of mononuclear cells. The expression of adhesion molecules on smooth muscle cells may be cell cycle-dependent, and influence smooth muscle cell proliferation and differentiation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Proteomic analysis of exosomes derived from human lymphoma cells.
701254 was used in western blot to characterize the proteomic content of lymphoma cell-derived exosomes.
|Yao Y,Wei W,Sun J,Chen L,Deng X,Ma L,Hao S||European journal of medical research (20:null)||2015|