CD65 antibody (FITC)

Catalog No. ABIN1112218

Product Details

Target: CD65

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: FITC

Application: Flow Cytometry (FACS), Immunofluorescence (IF)

Characteristics: Monoclonal Mouse Anti-Human CD65 FITC is recommended for use in flow cytometry for identification of human granulocytes.

Immunogen: HL60 promyelocytic cell line isolated from human acute myelogenous leukemia

Clone: 88H7

Isotype: IgG1

Application Details

Application Notes: It is recommended for use in flow cytometry. This reagent is effective for direct immunofluorescence staining of human tissue for flow cytometric analysis using 20 µl/10^6 cells.

Comment:

Fluorescein isothiocyanate.

Sample Collection: 1. Transfer 100 µl of anticoagulated (EDTA) blood to a 12 x 75 mm polystyrene test tube (10^6 cells). 2. Add 20 µl of CD65 FITC and mix gently with a vortex mixer. The 20 µl is a guideline only, the optimal volume should be determined by the individual laboratory. 3. The recommended negative control is a non-reactive FITC-conjugated antibody of the same isotype. 4. Incubate in the dark at room temperature at 4°C for 30 minutes or at room temperature (20-25 °C) for 15 minutes. 5. Add 1,5 ml of Lysing Solution to each sample and mix gently with a vortex mixer. Incubate for 10 minutes at room temperature in the dark. 6. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 7. Add 2 ml 0.01 mol/l PBS (It betters that it containing 2% bovine serum albumin) and resuspend the cells by using a vortex mixer. 8. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 9. Resuspend pellet in an appropriate fluid for flow cytometry, e.g. 0.3 ml PBS. The PBS should contain 1% paraformaldehyde (fixative) if samples are not analysed the same day. 10. Analyse on a flow cytometer or store at 2-8 °C in the dark until analysis. Samples can be run up to 24 hours after lysis. Fluorescence profiles of normal human peripheral blood Lymphocytes (blue histogram), Monocytes (red histogram) and Granulocytes (yellow histogram). Surface staining of normal human peripheral blood cells with Anti-Human CD65 FITC (cat. 65F-100T).

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: The conjugate is provided in liquid form in buffer PBS 20 mM, 1% BSA and 0,09 % Sodium azide.

Preservative: Sodium azide

Precaution of Use: 1. The device is not intended for clinical use including diagnosis, prognosis, and monitoring of a disease state, and it must not be used in conjunction with patient records or treatment. 2. This product contains Sodium azide (NaN3), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, Sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used.

Storage: 4 °C

Target Details

Target: CD65

Background: The antigen is expressed on myeloid cells and monocytes. CD65 is a fucosylated carbohydrate antigen on the cell surface. It is formed by fucosylation of the inner lactosamine unit of alpha-2,3-sialylated polylactosamine by alpha-1,3-fucosyltransferase (Niemela et al, 1998, Kono et al, 1997)