CD15 antibody

Catalog No. ABIN1112066

Product Details anti-CD15 Antibody

Target: CD15 (FUT4) (Fucosyltransferase 4 (Alpha (1,3) Fucosyltransferase, Myeloid-Specific) (FUT4))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD15 antibody is un-conjugated

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

Characteristics: Monoclonal Mouse Anti-Human CD15 PU, is recommended for use in flow cytometry for identification of neutrophils, monoblastoid precursor cells of the myeloid lineage, eosinophils, and some monocytes, but not on basophils and lymphocytes. The monoclonal antibody is provided in liquid form in buffer containing 1% bovine serum albumin (BSA) and 0,09% NaN3, pH 7.2.

Clone: MCS-1

Isotype: IgG3

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:

PURIFIED (pass through 0, 22 µm filter).

Sample Collection: 1. The starting material is peripheral blood (PB) or another sample of cells in suspension. This is permanently anticoagulated with EDTA (until the time of processing it should be stored at 4 °C). If processing is to be carried out within in a few hours, it may be more appropriate not to subject the cells to changes in temperature and leave the samples at room temperature. 2. Before starting the technique, a small sample is taken and read on a haematological counter to obtain the haemogram and determine the number of leukocytes. 3. Generally, 100 µL of PB are taken when the number of leukocytes is 10 x 10^3 cells/µL and 200/ µL when the number of leukocytes is equal to 5 x 10^-3 cells/ µL. 4. 1 µg of the McAb is pipetted into each tube and the tubes are incubated for 15 min at room temperature in the darkness. 5. A washing is made with centrifugation at 2000 rpm for 5 min with 5 ml of PBS in order to remove the McAb not bound to its antigen. 6. Add 10 µL of mouse anti-IgG antibody conjugated with some fluorochrome (Ref. Code No. GOATPOLYFITCANTIMOUIGG3) is added and the mixture is incubated at room temperature for 15 min in the darkness. The absence of light is necessary so that the fluorochrome will not deteriorate since it shows a high degree of photoinstability. 7. After the incubation period, an erythrocyte-lysing solution is added at the amount recommended by the manufacturer and the mixture is incubated at room temperature in the darkness (the blood should be well mixed with the lysing solution). 8. The tubes are centrifuged at 2000 rpm for 5 minutes. The supernatant is removed with a Pasteur pipette or with a vacuum pump. 9. The cell pellet is resuspended and a final wash is made with 3-5 ml of PBS at 2000 rpm for 5 min. 10. After removing the supernatant and resuspending the cell pellet, some 300 µL of PBS is added and the readings on the flow cytometer are recorded. 11. 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.

Restrictions: For Research Use only

Handling

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 for CD15

Target: CD15 (FUT4) (Fucosyltransferase 4 (Alpha (1,3) Fucosyltransferase, Myeloid-Specific) (FUT4))

Alternative Name: CD15 (FUT4 Products)

Synonyms: AI451562 antibody, CD15 antibody, FAL antibody, FucT-IV antibody, LeX antibody, SSEA-1 antibody, Ssea1 antibody, ELFT antibody, FCT3A antibody, FUC-TIV antibody, FUTIV antibody, Fuct antibody, fucosyltransferase 4 antibody, Fut4 antibody, FUT4 antibody

Background: The CD15 antigen is the X-hapten on lacto-N-fucose pentaosyl lll, which is expressed on circulating granulocytes or tissue granulocytes as well as neutrophils and eosinophils. The CD15 antibody recognizes the carbohydrate structure 3-fucosyl-N-acetyl-lactosamine. It specifically labels neutrophils, eosinophils and monoblastoid precursor cells of the myeloid lineage.