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Invitrogen™ CD177 Monoclonal Antibody (MEM-166 (MEM166)), eFluor™ 660, eBioscience™
Description
The MEM-166 monoclonal antibody reacts with human CD177, a 58-64 kDa GPI-anchored glycoprotein. The human CD177 locus codes for at least two alleles, NB1 and PRV-1, which differ by only 4 amino acids. CD177 is expressed on granulocytes, early erythroblasts, megakaryocytes, promyelocytes and myelocytes. CD177 mRNA levels are elevated in several different myeloproliferative disorders including polycythemia vera, and antibodies to CD177 are involved in neonatal alloimmune neutropenia, autoimmune drug induced neutropenia, and graft failure following marrow transplantation. Applications Reported: This MEM-166 (MEM166) antibody has been reported for use in flow cytometric analysis. Applications Tested: This MEM-166 (MEM166) antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (0.015 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. eFluor 660 is a replacement for Alexa Fluor 647. eFluor 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 μm post-manufacturing filtered.
Specifications
Specifications
| Antigen | CD177 |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | MEM-166 (MEM166) |
| Concentration | 5 μL/Test |
| Conjugate | eFluor 660 |
| Formulation | PBS with BSA and 0.09% sodium azide; pH 7.2 |
| Gene | CD177 |
| Gene Accession No. | Q8N6Q3 |
| Gene Alias | 1190003K14Rik; CD177; CD177 antigen; CD177 molecule; cell surface receptor; HNA2A; HNA-2a; Human neutrophil alloantigen 2a; NB1; NB1 glycoprotein; NB1 GP; Pdp3; Polycythemia rubra vera protein 1; PRV1; PRV-1; RGD1562941; UNQ595/PRO1181 |
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Frequently Asked Questions (FAQs)
Our options will depend on the samples you are analyzing.
If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.
For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.
Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).
Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.
The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.
As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.
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