Staining of C57Bl/6 bone marrow cells with eFluor® 450 anti-mo/hu B220 (RA3-6B2) (cat. 48-0452) and eFluor® 650NC Rat IgG2a Isotype Control (cat. 95-4321) (left) or eFluor® 650NC anti-mouse IgM (right). Total viable cells were used for analysis.
Contents: eFluor® 650NC anti-mouse IgM Catalog Number: 95-5790 Concentration: 5 µl Formulation: aqueous buffer, 0.09% sodium azide, contains stabilizer if necessary Storage Conditions: Store at 2-8°C. DO NOT FREEZE. LIGHT-SENSITIVE MATERIAL. Clone: II/41 Host/Isotype: Rat IgG2a, κ
Product Options
Cat. No.
Size
Price
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95-5790-41
25 tests
95-5790-42
100 tests
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*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.
Description
The II/41 monoclonal antibody reacts with the μ heavy chain of mouse IgM. It does not react with other classes of mouse immunoglobulin including IgD, IgG or IgA. IgM is expressed intracellularly, during early stages of B lymphopoiesis, and then on the surface of more mature B cells in the bone marrow and peripheral B cells. Fluorochrome conjugated II/41 can be used as a detection secondary for mouse IgM.
Applications Reported
For research use only, not for diagnostic or therapeutic use. This II/41 antibody has been reported for use in flow cytometric analysis.
Applications Tested
This II/41 antibody has been pre-titrated and tested by flow cytometric analysis of mouse bone marrow cells using the eFluor® NC Flow Cytometry Staining Buffer (cat. 00-3222). This can be used at 5 μl per million cells in a 100 μl total staining volume.
The isotype control eFluor® 650NC rat IgG2a (cat. 95-4321) should be used at 5 ul/test.
Laser/Filter Recommendation: When using eFluor 650NC, we recommend excitation with the 405nm violet laser with an appropriate filter set, such as the 630 LP dichroic mirror with the 660/40 bandpass filter. The eFluor 650NC can be minimally excited off of the 633 nm laser, and because its peak emission is 650nm, it will require some compensation out of the APC detector. Please contact eBioscience Technical Support for more information.
Buffer Recommendation: Comparison of eFluor® NC conjugated antibody staining in different buffers has demonstrated that optimal performance is seen with the eFluor® NC Flow Cytometry Staining Buffer (cat. 00-3222). For a comparison of staining with different buffers, click here.
Click here for answers to questions about fixation and other FAQs in the eFluor® Nanocrystal Frequently Asked Questions.
References
Laszlo G, Hathcock KS, Dickler HB, Hodes RJ. 1993. Characterization of a novel cell-surface molecule expressed on subpopulations of activated T and B cells. J Immunol. 150(12):5252-62.
