Description: The IM7 monoclonal antibody reacts with all isoforms of mouse CD44 (Pgp-1). CD44 is expressed by hematopoietic and non-hematopoietic cells. Bone marrow myeloid cells and memory T cells highly express this antigen and peripheral B and T cells can upregulate the expression of CD44. CD44 functions as an adhesion molecule through its binding to hyaluronate, an extracellular matrix component.
Applications Reported: This IM7 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This IM7 antibody has been tested by flow cytometric analysis of mouse bone marrow cells and splenocyte suspensions. 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 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 Complete Staining Buffer (Product # SB-4401) 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 # 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 # A10497).
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 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
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
CD44 cell surface antigen is a 100 kDa type 1 transmembrane glycoprotein widely expressed on human leucocytes, white matter of the brain and by some epithelial cells of the intestine and breast. Several isoforms of CD44 exist, including the predominant CD44H isoform detected in many normal tissues. CD44 is a receptor for hyaluronic acid (HA) and is involved in cell-cell interactions, cell adhesion and migration. CD44 also participates in a wide variety of cellular functions including lymphocyte activation, recirculation and homing. CD44 expression may be up-regulated upon some carcinomas, and it has been speculated that this may be related to metastatic potential. CD44 is expressed by hematopoietic, non-hematopoietic cells, epithelial tissues, and to filopodia in cultured keratinocytes. Further, bone marrow myeloid cells and memory T cells express CD44 at high levels, and peripheral B and T cells can upregulate the expression of CD44 in response to certain stimulatory events. Transcripts for the CD44 gene undergo complex alternative splicing that results in many functionally distinct isoforms, however, the full-length nature of some of these variants have not been determined. Alternative splicing is the basis for the structural and functional diversity of the CD44 protein. Diseases associated with CD44 dysfunction include superficial keratitis and lichen sclerosus. CD44 also may be related to tumor metastasis formation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: CD44; CD44 antigen; CD44 molecule; CDw44; cell surface glycoprotein CD44; chondroitin sulfate proteoglycan 8; ECMR-III; ECMRIII; Epican; Extracellular matrix receptor III; GP90 lymphocyte homing/adhesion receptor; hematopoietic cell E- and L-selectin ligand; Heparan sulfate proteoglycan; Hermes antigen; homing function and Indian blood group system; HUTCH-I; hyaluronan receptor; Hyaluronate receptor; Ly-24; Lymphocyte antigen 24; PGP I; PGP-1; PGP-I; PGPI; Phagocytic glycoprotein 1; Phagocytic glycoprotein I; sCD 44; sCD44; soluble CD 44; soluble CD44
Gene Aliases: AU023126; AW121933; AW146109; CD44; CDW44; CSPG8; ECMR-III; HCELL; HERMES; HUTCH-I; IN; LHR; Ly-24; MC56; MDU2; MDU3; MIC4; Pgp-1; Pgp1
Molecular Function: transmembrane signal receptor