Description: The N418 monoclonal antibody reacts with mouse CD11c, the integrin alphaX. CD11c non-covalently associates with beta2 integrin to form the CD11c/CD18 heterodimer. CD11c is expressed by dendritic cells, a subset of Intestinal Intraepithelial Lymphocytes (IEL) and some activated T cells. CD11c/CD18 binds to CD54, iC3b and fibrinogen and plays a role in leukocyte adhesive interactions. N418 binds to CD11c on splenic dendritic cells in the T-dependent areas of mouse spleen and precipitates a 150, 90 kDa heterodimer. Applications Reported: This N418 antibody has been reported for use in flow cytometric analysis. Applications Tested: This N418 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.5 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Super Bright 780 can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter, or equivalent. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Staining Buffer (Product No.SB-4400) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product No. 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product No. A10497). Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product No. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser CD11b (integrin alpha-M, ITGAM, integrin alpha-X, ITGAX) is a 165 kDa adhesion molecule that associates non-covalently with integrin beta-2 (CD18). The CD11b/CD18 heterodimeric complex is also known as integrin alpha-M beta-2, Mac-1, and CR3 (complement receptor 3). CD11b is expressed on the surface of monocytes/macrophages, granulocytes, activated lymphocytes, a subset of NK cells, a subset of dendritic cells, and microglia in the brain. CD11b/CD18 functions as the receptor for ICAM-1 (CD54), ICAM-2 (CD102), ICAM-4 (CD242), CD14, CD50, CD23, heparin, iC3b, fibrinogen, and Factor X -these adhesions are critical for cell-cell and cell-matrix interactions. CD11b is expressed on 8% of spleen cells, 44% of bone marrow cells, and less than 1% of thymocytes, and is commonly used as a microglial marker in nervous tissue. The expression of CD11b increases during monocyte maturation and expression levels vary on tissue macrophages. Further, peritoneal macrophages are reported to express higher levels of CD11b than splenic macrophages. Diseases associated with CD11b dysfunction include systemic lupus erythematosus 6 and ITGAM-related susceptibility to systemic lupus erythematosus.
|4° C, store in dark, DO NOT FREEZE!|
|Super Bright 780|
|PBS with BSA and 0.09% sodium azide; pH 7.2|