Mouse anti-Human CD65s Antibody | anti-CD65s antibody
Mouse anti CD65s
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.
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
A variety of studies have demonstrated the usefulness and reliability of VIM2 as a marker molecule for the classification of acute leukemias. Recently, the signal transducing capacity of VIM2 bearing surface molecules has been demonstrated.
The VIM2 antibody permits the identification and enumeration of normal and leukemic cell populations expressing the VIM2 antigen present in human biological samples (blood, bone marrow and others) using flow cytometry. Furthermore, VIM2 mAb is suitable for the elimination of myeloid cells from complex cell mixtures as well as for functional studies. (Lund-Johansen et al.) Results must be put within the context of other tests as well as the clinical history of the patient by a certified professional before
final interpretation. Analysis performed with this antibody should be paralleled by positive and negative controls.
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).
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Product Notes
The CD65s (Catalog #AAA570356) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Mouse anti CD65s reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's CD65s can be used in a range of immunoassay formats including, but not limited to, Direct Immunofluorescence (IF), Indirect Immunofluorescence (IF), Flow Cytometry (FC/FACS). 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. Researchers should empirically determine the suitability of the CD65s for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CD65s, Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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