CD22 anticorps (PE)

N° du produit ABIN1741592

L’anticorps Souris Monoclonal anti-CD22 a été validé pour FACS et IFA. Il convient pour détecter CD22 dans des échantillons de Humain. Il y a 14+ publications disponibles.

Aperçu rapide pour CD22 anticorps (PE) (ABIN1741592)

Antigène: CD22 (CD22 Molecule (CD22))

Reactivité: Humain

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp CD22 est conjugé à/à la PE

Application: Flow Cytometry (FACS), Indirect Immunofluorescence Assay (IFA)

Clone: RFB4

Détail du produit anti-CD22 anticorps (PE)

Fonction: This product is optimised for use with FIX&PERM®.

Marque: FIX&PERM®

Specificité: The CD22 mAb (clone RFB4) recognizes surface CD22 expressed by mature peripheral B-cells and cytoplasmatic CD22 expressed by precursor B-cells. The sensitivity of RFB4 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 fluorescence intensity).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, 50 µL of leukocytes containing 10^7 cells/mL are stained with 20 µL 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 3-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.

Attributs du produit: Mildly fixes cells, preserving their flow cytometric scatter characteristics
Allows simultaneous characterisation of both intracellular and cell surface markers
Rapid technique - whole procedure can be carried out in less than one hour, ready
for immediate analysis or storage for 24 hours
Stringent QC procedures - the quality of each lot is determined using well-defined
blood samples and subsequent comparison of scatter characteristics of obtained
leukocyte populations, ensuring consistent and reliable results lot after lot
A range of intracellular antibodies with optimised protocols for use with FIX&PERM®
FIX&PERM® is a simple procedure making use of two reagents. Reagent A gently fixes cells, while Reagent B permeabilizes them. The specific formulations reduce background and allow simultaneous addition of permeabilization medium and fluorochrome labelled antibodies, allowing staining of intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulins, etc.

Purification: Purified by Affinity Chromatography

Isotype: IgG1

Information d'application

Indications d'application: Staining Procedure for Surface CD22: Direct Immunofluorescence (Staining Procedure) fluorochrome labeled antibodies are designed for use with either whole blood or isolated mononuclear cell (MNC) preparationsProposed staining procedure for whole blood in short: - For each sample add 50 µL of EDTA anti-coagulated blood to a 3-5 mL tube - Add 20 µL of the appropriate monoclonal antibody conjugate - Incubate the tube for 15 minutes at 4°C or at room temperature in the dark - Add 100 µL -LYSE (ABIN1741580) 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° C in the dark and analyze within 24 hoursFor \No-Wash\ Proposed staining procedure for MNC in short: - Carefully add 20 µL antibody conjugate and 50-100 µL MNC to the bottom of a tube - Vortex at low speed for 1-2 seconds - Incubate for 15-30 minutes at 2-8°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°C in the dark. - Analyze fixed cells within 24 hours Indirect Immunofluorescence (Staining Procedure) - Mix 20 µL purified antibody with 50 µL whole blood or MNC suspension - Incubate for 15 minutes at 2-8°C - Wash cells with phosphate buffered saline (PBS)- Add to cell pellet 20 µL of affinity purified, fluorochrome labeled F(ab')2 anti mouse Ig antibodies - Incubate for 15 minutes at 2-8°C - Wash cells with phosphate buffered saline (PBS) or proceed as described for direct stainingStaining Procedure for Cytoplasmatic CD22: Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM® (ABIN1741575) intracellular CD22 can be easily stained in cell suspensions. - For each sample to be analyzed add 50 µL of whole blood, bone marrow or mononuclear cell suspension in a 5 mL tube- Add 100 µL of Reagent A (Fixation Medium, stored and used at room temperature) - Incubate for 15 minutes at room temperature - Add 5 mL phosphate buffered saline and centrifuge cells for 5 minutes at 300 g - Remove supernatant and add to cell pellet 100 µL Reagent B (Permeabilization Medium) and 20 µL of the CD22 monoclonal antibody conjugate - Vortex at low speed for 1-2 seconds - Incubate for 15 minutes at room temperature - Wash cells with phosphate buffered saline as described above - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 mL 1.0 % formaldehyde and store them at 2- 8°C in the dark. - Analyze fixed cells within 24 hours.

Commentaires:

The epitope recognized by antibody RFB4 is a 135 kDa type I integral membrane protein expressed by human B-cells. Precursor B-cells are surface-CD22 negative, but cytoplasmic CD22 positive. Mature B-lymphocytes express CD22 also on their surface.The RFB4 antibody permits the identification and enumeration of human B-cells using flow cytometry. Results must be put within the context of other diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation. Analyses performed with this antibody should be paralleled by positive and negative controls.

Procédure de l'essai:

Procedure: For each sample to be analysed use 50 µL of whole blood, bone marrow or mononuclear cell suspension in a 5 mL tube
Add 100 µL of Reagent A (Fixation Medium, stored and used at room temperature)
Incubate for 15 minutes at room temperature
Add 5 mL phosphate buffered saline and centrifuge cells for 5 minutes at 300 g
Remove supernatant and add to cell pellet 100 µL Reagent B (Permeabilization
Medium) and 20 µL of the appropriate monoclonal antibody conjugate
Vortex at low speed for 1-2 seconds
Incubate for 15 minutes at room temperature
Wash cells with phosphate buffered saline as described above
Remove supernatant and re-suspend cells in sheath fluid for immediate analysis or re-suspend cells in 0.5 mL 1.0 % formaldehyde and store at 2-8°C in the dark
Analyse fixed cells within 24 hours

Restrictions: For Research Use only

Stockage

Format: Liquid

Buffer: PBS pH 7.2, 1 % BSA, 0.05 % sodium azide

Agent conservateur: Sodium azide

Précaution d'utilisation: This reagent contains sodium azide. To avoid the development of hazardous conditions, reagents containing azide should be diluted in running water prior to be discarded. Similar to the work with other biological products, proper handling procedures are recommended.

Conseil sur la manipulation: Do not freeze and protect from prolonged exposure to light.

Stock: 4 °C

Stockage commentaire: These reagents should be stored at 2-8°C 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.

Références pour anticorps anti-CD22 (PE) (ABIN1741592)

Braylan, Orfao, Borowitz, Davis: "Optimal number of reagents required to evaluate hematolymphoid neoplasias: results of an international consensus meeting." dans: Cytometry, Vol. 46, Issue 1, pp. 23-7, (2001) (PubMed).

Lanza, Latorraca, Moretti, Castagnari, Ferrari, Castoldi: "Comparative analysis of different permeabilization methods for the flow cytometry measurement of cytoplasmic myeloperoxidase and lysozyme in normal and leukemic cells." dans: Cytometry, Vol. 30, Issue 3, pp. 134-44, (1997) (PubMed).

Groeneveld, te Marvelde, van den Beemd, Hooijkaas, van Dongen: "Flow cytometric detection of intracellular antigens for immunophenotyping of normal and malignant leukocytes." dans: Leukemia, Vol. 10, Issue 8, pp. 1383-9, (1996) (PubMed).

Catovsky, Matutes, Buccheri, Shetty, Hanslip, Yoshida, Morilla: "A classification of acute leukaemia for the 1990s." dans: Annals of hematology, Vol. 62, Issue 1, pp. 16-21, (1991) (PubMed).

Stamenkovic, Seed: "The B-cell antigen CD22 mediates monocyte and erythrocyte adhesion." dans: Nature, Vol. 345, Issue 6270, pp. 74-7, (1990) (PubMed).

Li, Shen, Ghetie, May, Till, Ghetie, Uhr, Janossy, Thorpe, Amlot: "The epitope specificity and tissue reactivity of four murine monoclonal anti-CD22 antibodies." dans: Cellular immunology, Vol. 118, Issue 1, pp. 85-99, (1989) (PubMed).

Janossy, Coustan-Smith, Campana: "The reliability of cytoplasmic CD3 and CD22 antigen expression in the immunodiagnosis of acute leukemia: a study of 500 cases." dans: Leukemia, Vol. 3, Issue 3, pp. 170-81, (1989) (PubMed).

Rani, De Oliveira, Catovsky: "Different expression of CD3 and CD22 in leukemic cells according to whether tested in suspension or fixed on slides." dans: Hematologic pathology, Vol. 2, Issue 2, pp. 73-8, (1989) (PubMed).

van der Schoot, von dem Borne, Tetteroo: "Characterization of myeloid leukemia by monoclonal antibodies, with an emphasis on antibodies against myeloperoxidase." dans: Acta haematologica, Vol. 78 Suppl 1, pp. 32-40, (1988) (PubMed).

Boué, Lebien: "Structural characterization of the human B lymphocyte-restricted differentiation antigen CD22. Comparison with CD21 (complement receptor type 2/Epstein-Barr virus receptor)." dans: Journal of immunology (Baltimore, Md. : 1950), Vol. 140, Issue 1, pp. 192-9, (1988) (PubMed).

Pezzutto, Rabinovitch, Dörken, Moldenhauer, Clark: "Role of the CD22 human B cell antigen in B cell triggering by anti-immunoglobulin." dans: Journal of immunology (Baltimore, Md. : 1950), Vol. 140, Issue 6, pp. 1791-5, (1988) (PubMed).

Mason, Stein, Gerdes, Pulford, Ralfkiaer, Falini, Erber, Micklem, Gatter: "Value of monoclonal anti-CD22 (p135) antibodies for the detection of normal and neoplastic B lymphoid cells." dans: Blood, Vol. 69, Issue 3, pp. 836-40, (1987) (PubMed).

Dörken, Pezzutto, Moldenhauer, Schwartz, Kiesel, Hunstein: "An immunoenzymatic staining assay (ISA) for the rapid screening of monoclonal antibodies detecting membrane and cytoplasmic antigens." dans: Journal of immunological methods, Vol. 88, Issue 1, pp. 129-36, (1986) (PubMed).

Dörken, Moldenhauer, Pezzutto, Schwartz, Feller, Kiesel, Nadler: "HD39 (B3), a B lineage-restricted antigen whose cell surface expression is limited to resting and activated human B lymphocytes." dans: Journal of immunology (Baltimore, Md. : 1950), Vol. 136, Issue 12, pp. 4470-9, (1986) (PubMed).

Détails sur CD22

Antigène: CD22 (CD22 Molecule (CD22))

Autre désignation: CD22

Poids moléculaire: 135 kDa