Description: The 61D3 monoclonal antibody reacts with human CD14, a 53-55 kDa GPI-linked glycoprotein. CD14 is expressed on monocytes, interfollicular macrophages and some dendritic cells. Complexes of LPS and LBP (LPS-Binding Protein) bind with high affinity to monocytes through the surface CD14. Applications Reported: This 61D3 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 61D3 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 μL (0.25 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. Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. 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.
|Baboon, Chimpanzee, Cynomolgus Monkey, Human, Rhesus Monkey|
|4° C, store in dark, DO NOT FREEZE!|
|Super Bright 780|
|PBS with BSA and 0.09% sodium azide; pH 7.2|