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CD65s Antibody – Mouse

For Research Use Only. Not for use in diagnostic procedures. Catalog #: MBS570356

The epitope recognized by antibody VIM2 is expressed by virtually all myeloid cells including normal and malignant granulocytes and monocytes. In normal myelopoiesis VIM2 can first be detected after the late CFU-GM stage. In acute myeloid leukemias (AMLs) in… More details

$500 / 0.2 mg +1 FREE 8GB USB
Product Documentation
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Sample Manual Insert View PDF
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Testing Data View PDF
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Certificate of Analysis (COA) View PDF
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QC Report View PDF
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Material Safety Data Sheet View PDF
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Technical Datasheet View PDF
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Protocol View PDF

Product Overview

Online Datasheet in PDF
Product Name
CD65s, Antibody
Full Product Name
Mouse anti CD65s
Product Gene Name
anti-CD65s antibody
Isotype
IgM
Clone Number
VIM2
Host
Mouse
Species Reactivity
Human
Form/Format
PBS pH 7.2, 1% BSA, 0.05% NaN3
Specificity
Antibody VIM2 reacts with a carbohydrate structure expressed by myeloid cells. The epitope recognized was shown by Macher et al. to involve a defined sialofucooligosaccharide sequence. Similar results were obtained by Kniep et al.. Together with other sialylated and fucosylated polylactosamines the carbohydrate structure recognized by VIM2 may play a critical role on the adhesion of granulocytes and monocytes to endothelium and platelets during inflammation and clotting.

The sensitivity of VIM2 mAb is determined by staining well-defined blood samples from representative donors with serial-fold mAb dilutions to obtain a titration curve that allows relating the mAb concentration to the percentage of stained cells and geometric MFI (mean fluorescenceintensity). For this purpose, a mAb-concentration range is selected to include both the saturation point (i.e. the mAb dilution expected to bind all epitopes on the target cell) and the detection threshold (i.e. the mAb dilution expected to represent the least amount of mAb needed to detect an identical percentage of cells). In practice, 50ul of leukocytes containing 10^7
cells/ml are stained with 20ul mAb of various dilutions to obtain a titration curve and to identify the saturation point and detection
threshold. The final concentration of the product is then adjusted to be at least three-fold above the detection threshold. In addition and to control lot-to-lot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.
Preparation and Storage
Monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody. For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8 degree C (DO NOT FREEZE!) and protected from prolonged exposure to light. Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended.
CE Mark
CE
Applications Tested/Suitable for anti-CD65s antibody
Direct Immunofluorescence (IF), Indirect Immunofluorescence (IF), Flow Cytometry (FC/FACS)
Application Notes for anti-CD65s antibody
Staining Procedure
Direct Immunofluorescence (Staining Procedure)
Fluorochrome labeled antibodies are designed for use with either whole blood or isolated mononuclear cell (MNC) preparations.

Proposed staining procedure for whole blood in short:
- For each sample add 50 ul of EDTA anti-coagulated blood to a 3-5 ml tube
- Add 20 ul of the appropriate monoclonal antibody conjugate
- Incubate the tube for 15 minutes at 4 degree C or at room temperature in the dark
- Add 100 ul NM-LYSE to each tube and incubate for 10 minutes at room temperature - Add 3-4 ml of destilled water and vortex, incubate for 5-10
minutes at room temperature
- Centrifuge tube for 5 minutes at 300 g - Aspirate supernatant and resuspend pellet in 0.3 ml of sheath fluid
- Analyze immediately or store samples at 2-8 degree C in the dark and analyze within 24 hours
Proposed staining procedure for MNC in short:
- Carefully add 20 ul antibody conjugate and 50-100 ul MNC to the bottom of a tube
- Vortex at low speed for 1-2 seconds
- Incubate for 15-30 minutes at 2-8 degree C or at room temperature
- Centrifuge tubes for 5 minutes at 300 g
- Remove supernatant, resuspend cells in 2-5 ml of phosphate buffered saline (PBS) and centrifuge cells again for 5 minutes at 300 g
- Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1 % formaldehyde and store them at 2-8 degree C in the dark. Analyze
fixed cells within 24 hours

Indirect Immunofluorescence (Staining Procedure)
- Mix 20 ul purified antibody with 50 ul whole blood or MNC suspension
- Incubate for 15 minutes at 2-8 degree C
- Wash cells with phosphate buffered saline (PBS)
- Add to cell pellet 20 ul of affinity purified, fluorochrome labeled F(ab')2 anti mouse Ig antibodies
- Incubate for 15 minutes at 2-8 degree C
- Wash cells with phosphate buffered saline (PBS) or proceed as described for direct staining

Product References and Citations for anti-CD65s antibody

O. Majdic et al., Int J Cancer 33, 617 (1984).
P. Bettelheim et al., Leuk Res 9, 1323 (1985).
C. Peschel et al., Exp Hematol 13, 1211 (1985).
K. Uemura et al., Biochim Biophys Acta 846, 26 (1985).
D. Lutz et al., Onkologie 9, 67 (1986).
R. Delwel, F. Bot, W. Knapp, B. Lowenberg, Bone Marrow Transplant 2, 149 (1987).
B. A. Macher, J. Buehler, P. Scudder, W. Knapp, T. Feizi, J Biol Chem 263, 10186 (1988).
U. Koller et al., Leukemia 3, 708 (1989)
B. A. Macher, J. H. Beckstead, Leuk Res 14, 119 (1990).
I. Schwarzinger et al., J Clin Oncol 8, 423 (1990).
J. B. Lowe et al., J Biol Chem 266, 17467 (1991).
T. A. Springer, L. A. Lasky, Nature 349, 196 (1991).
F. Lund-Johansen et al., J Immunol 148, 3221 (1992).
F. M. Fink et al., Med Pediatr Oncol 21, 340 (1993).
F. Lund-Johansen et al., Eur J Immunol 23, 2782 (1993).
J. Stockl et al., J Leukoc Biol 53, 541 (1993).
W. Knapp, H. Strobl, O. Majdic, Cytometry 18, 187 (1994).
G. M. Brown, T. N. Huckerby, B. L. Abram, I. A. Nieduszynski, Biochem J 319 (Pt 1), 137 (1996).
J. L. Clarke, W. Watkins, J Biol Chem 271, 10317 (1996).
R. N. Knibbs et al., J Cell Biol 133, 911 (1996).
B. Kniep et al., J Biochem (Tokyo) 119, 456 (1996).
A. J. Wagers, L. M. Stoolman, R. Kannagi, R. Craig, G. S. Kansas, J Immunol 159, 1917 (1997).
M. Noguchi, N. Sato, H. Sugimori, K. Mori, K. Oshimi, Leuk Res 25, 847 (2001).
W. M. Watkins, J. L. Clarke, Adv Exp Med Biol 491, 231 (2001).

Other Notes

Small volumes of anti-CD65s antibody vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

Searchable Terms for CD65s purchase

MBS570356 is an antibody targeting CD65s. Also known as Mouse anti CD65s. Reactive in Human. Host: Mouse.
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