Spark Red™ 718 anti-mouse CD11a (Flexi-Fluor™) Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

C57BL/6 mouse splenic secondary cytotoxic T cells

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide

Preparation

The antibody was purified by affinity chromatography and conjugated with Spark Red™ 718 under optimal conditions.

Concentration

0.2 mg/mL

Storage & Handling

The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.

Application

FC

Recommended Usage

Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.

For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 10⁷ cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.

Excitation Laser

Red Laser (633 nm)

Application Notes

The M17/4 antibody can block CD11a-mediated cellular adhesion. Additional reported applications of this antibody (for the relevant formats) include: immunoprecipitation^1,2, in vitro blocking of cell-cell adhesion^1,2 and FOXP3 expression⁵, and immunohistochemical staining of acetone-fixed frozen sections³. The M17/4 antibody does not block the binding of 2D7 antibody (Cat. No. 101002) to CD11a. The LEAF™ purified antibody (Endotoxin <0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 101110). For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 101118) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin <0.01 EU/µg).

Additional Product Notes

For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.

Application References

(PubMed link indicates BioLegend citation)

1. Sanchez-Madrid F, et al. 1982. Cell Immunol. 73:1. (IP, Block)
2. Kuhlman P, et al. 1991. J. Immunol. 146:1773. (IP, Block)
3. Mizgerd JP, et al. 1997. J. Exp. Med. 186:1357. (IHC)
4. Hailman E and Allen PM. 2005. J. Immunol. 175:4847. (FC)
5. Verhagen J and Wraith DC. 2014. J. Immunol. Methods. S0022. (Block) PubMed

RRID

AB_3106342 (BioLegend Cat. No. 285087)

Antigen Details

Structure

Integrin family, associates with integrin β2 (CD18), 180 kD

Distribution

Lymphocytes, granulocytes, monocytes, macrophages

Function

Cellular adhesion

Ligand/Receptor

ICAM-1 (CD54), ICAM-2 (CD102), ICAM-3 (CD50)

Cell Type

Granulocytes, Lymphocytes, Macrophages, Monocytes, Tregs

Biology Area

Cell Adhesion, Cell Biology, Costimulatory Molecules, Immunology, Innate Immunity, Neuroinflammation, Neuroscience

Molecular Family

Adhesion Molecules, CD Molecules

Antigen References

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Springer TA. 1994. Cell 76:301.
3. Lub M, et al. 1995. Immunol. Today 16:479.

Gene ID

16408 View all products for this Gene ID

UniProt

View information about CD11a on UniProt.org

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Other Formats