KAPPA, CD19 antibody

Catalog No. ABIN1112413

Product Details

Target: KAPPA, CD19

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

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

Characteristics: Mouse monoclonal Anti-Human KAPPA FITC/CD19 PE, is recommended for use in flow cytometry.

Clone: Nam75-5F3-A3-B1

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.

Sample Preparation:

1. Transfer 100 µl of anticoagulated (EDTA) blood to a 12 x 75 mm polystyrene test tube. 2. Add 2 ml 0.01 mol/l PBS ( It better that it containing 2% bovine serum albumin) and resuspend the cells by using a vortex mixer. 3. Incubate for 10 minutes at 37°C. 4. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 5. Add 2 ml 0.01 mol/l PBS ( It better that it containing 2% bovine serum albumin) and resuspend the cells by using a vortex mixer. 6. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 7. Add 20 µl of LAMBDA/CD19 and mix gently with a vortex mixer. The 20 µl is a guideline only, the optimal volume should be determined by the individual laboratory. 8. The recommended negative control is a non-reactive FITC-conjugated antibody of the same isotype and a non-reactive PE-conjugated antibody of the same isotype . 9. Incubate in the dark at room temperature at 4°C for 30 minutes or at room temperature (20-25 °C) for 15 minutes. 10. 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. 11. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 12. Add 2 ml 0.01 mol/l PBS ( It better that it containing 2% bovine serum albumin) and resuspend the cells by using a vortex mixer. 13. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 14. 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. 15. 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

Target: KAPPA, CD19

Background: Anti- Kappa Light Chains reacts with free kappa chains as well as kappa chains in intact immunoglobulin molecules. The specificity has been ascertained as described below. Flow cytometry: When Anti- Kappa Light Chains is applied as described in the staining procedure in combination with Anti- CD19/ RPE, on lysed human whole blood, a specific staining of a part of the CD19- positive B lymphocytes is seen corresponding to the expected range of kappa light chain expression. Clinical applications: Flow cytometric analysis of single cell suspensions from formalin- fixed, paraffin- embedded tissue samples demonstrated that Anti- Kappa Light Chains labels reactive hyperplastic lymph nodes (10/ 10 cases). non- Hodgkin-s lymphomas, the antibody labeled 4/ 10 cases. The remaining cases were positive for lambda light chains (5/ 10 cases) or showed no expression of light chains (1/ 10 cases). Flow cytometric analysis of peripheral blood lymphocytes demonstrated that Anti- Kappa Light Chains labels a proportion of B- cell chronic lymphocytic leukaemias. Thus, in one study of 121 cases , 37 were positive for kappa. another study of 165 cases (3), 97 were positive for kappa. Anti- CD19, labels human B cells in peripheral blood, bone marrow and other tissues. Additionally, B- cell lymphoproliferative disorders gave positive reactions with the antibody, i. e. acute lymphoblastic leukaemia, chronic lymphocytic leukaemia, hairy cell leukaemia, lymphoblastic lymphoma (Burkitt type), centroblastic/ centrocytic lymphoma, and diffuse non- Hodgkin-s lymphoma. The antibody was shown to be unreactive with other cells in the human haema-topoietic system and did not react with non- haematopoietic cells, e. g. kidney, liver, breast or lung tissues.