CD68 anticorps (FITC)

N° du produit ABIN1741593

Cet anticorps anti-CD68 est un anticorps Souris Monoclonal détectant CD68 dans FACS. Adapté pour Humain. Ce Primary Antibody a été cité dans 12+ publications.

Aperçu rapide pour CD68 anticorps (FITC) (ABIN1741593)

Antigène: CD68 (CD68 Molecule (CD68))

Reactivité: Humain

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp CD68 est conjugé à/à la FITC

Application: Flow Cytometry (FACS)

Clone: Ki-M7

Détail du produit anti-CD68 anticorps (FITC)

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

Specificité: The CD68 mAb (clone Ki-M7) reacts with human macrosialin. The sensitivity of CD68 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 identical percentage of cells). In practice, 50µL of leukocytes containing 10^7 cells/mL are stained with 20µL 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: Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM- (ABIN1741575) intracellular CD68 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 100µl Reagent B (Permeabilization Medium) and 20 µL of the CD68 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:

CD68 (macrosialin) is a type-one membrane protein with significant sequence homology to a family of lysosomal-associated glycoproteins (lamp). CD68 molecules are predominantly located intracellularily in cytoplasmic granules but can also be detected in smaller amounts at the cell surface. Particular strong intracellular expression is observed for monocytes and macrophages. In addition, Langerhans cells as well as plasmacytoid dendritic cells express clear-cut levels of CD68. Low-level reactivity is also observed with a subset of B lymphocytes and activated T lymphocytes. Oxidized low-density lipoprotein is a ligand for CD68. The CD68 mAb permits the identification and enumeration of monocytes, Langerhans cells and plasmacytoid dendritic cells (in combination with CD4/CD56 staining) in normal and malignant human blood and bone marrow samples 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-CD68 (FITC) (ABIN1741593)

Strobl, Knapp: "Myeloid cell-associated lysosomal proteins as flow cytometry markers for leukocyte lineage classification." dans: Journal of biological regulators and homeostatic agents, Vol. 18, Issue 3-4, pp. 335-9, (2005) (PubMed).

Feuillard, Jacob, Valensi, Maynadié, Gressin, Chaperot, Arnoulet, Brignole-Baudouin, Drénou, Duchayne, Falkenrodt, Garand, Homolle, Husson, Kuhlein, Le Calvez, Sainty, Sotto, Trimoreau, Béné: "Clinical and biologic features of CD4(+)CD56(+) malignancies." dans: Blood, Vol. 99, Issue 5, pp. 1556-63, (2002) (PubMed).

Sadovnikova, Parovichnikova, Savchenko, Zabotina, Stauss: "The CD68 protein as a potential target for leukaemia-reactive CTL." dans: Leukemia, Vol. 16, Issue 10, pp. 2019-26, (2002) (PubMed).

Strobl, Scheinecker, Riedl, Csmarits, Bello-Fernandez, Pickl, Majdic, Knapp: "Identification of CD68+lin- peripheral blood cells with dendritic precursor characteristics." dans: Journal of immunology (Baltimore, Md. : 1950), Vol. 161, Issue 2, pp. 740-8, (1998) (PubMed).

Ramprasad, Terpstra, Kondratenko, Quehenberger, Steinberg: "Cell surface expression of mouse macrosialin and human CD68 and their role as macrophage receptors for oxidized low density lipoprotein." dans: Proceedings of the National Academy of Sciences of the United States of America, Vol. 93, Issue 25, pp. 14833-8, (1997) (PubMed).

Scheinecker, Strobl, Fritsch, Csmarits, Krieger, Majdic, Knapp: "Granulomonocyte-associated lysosomal protein expression during in vitro expansion and differentiation of CD34+ hematopoietic progenitor cells." dans: Blood, Vol. 86, Issue 11, pp. 4115-23, (1996) (PubMed).

Ramprasad, Fischer, Witztum, Sambrano, Quehenberger, Steinberg et al.: "The 94- to 97-kDa mouse macrophage membrane protein that recognizes oxidized low density lipoprotein and phosphatidylserine-rich liposomes is identical to macrosialin, the mouse homologue of human ..." dans: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, Issue 21, pp. 9580-4, (1995) (PubMed).

Knapp, Strobl, Majdic: "Flow cytometric analysis of cell-surface and intracellular antigens in leukemia diagnosis." dans: Cytometry, Vol. 18, Issue 4, pp. 187-98, (1995) (PubMed).

Strobl, Scheinecker, Csmarits, Majdic, Knapp: "Flow cytometric analysis of intracellular CD68 molecule expression in normal and malignant haemopoiesis." dans: British journal of haematology, Vol. 90, Issue 4, pp. 774-82, (1995) (PubMed).

Remaley, Ugorski, Wu, Litzky, Burger, Moore, Fukuda, Spitalnik: "Expression of human glycophorin A in wild type and glycosylation-deficient Chinese hamster ovary cells. Role of N- and O-linked glycosylation in cell surface expression." dans: The Journal of biological chemistry, Vol. 266, Issue 35, pp. 24176-83, (1992) (PubMed).

Rabinowitz, Gordon: "Macrosialin, a macrophage-restricted membrane sialoprotein differentially glycosylated in response to inflammatory stimuli." dans: The Journal of experimental medicine, Vol. 174, Issue 4, pp. 827-36, (1991) (PubMed).

Davey, Cordell, Erber, Pulford, Gatter, Mason: "Monoclonal antibody (Y1/82A) with specificity towards peripheral blood monocytes and tissue macrophages." dans: Journal of clinical pathology, Vol. 41, Issue 7, pp. 753-8, (1988) (PubMed).

Détails sur CD68

Antigène: CD68 (CD68 Molecule (CD68))

Autre désignation: CD68