Lactoferrin anticorps (PE)

N° du produit ABIN1741598

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

Aperçu rapide pour Lactoferrin anticorps (PE) (ABIN1741598)

Antigène: Lactoferrin (LTF) (Lactotransferrin (LTF))

Reactivité: Humain

Hôte: Souris

Clonalité: Monoclonal

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

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

Clone: 4C5

Détail du produit anti-Lactoferrin anticorps (PE)

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

Marque: FIX&PERM®

Specificité: The LF mAb (clone 4C5) recognizes Lactoferrin stored within secondary granules of postmitotic granulocyte-committed cells. The sensitivity of 4C5 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 10^7cells/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: Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM® (ABIN1741575) intracellular Lactoferrin 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 Lactoferrin 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. Conventional Staining for Microscopic Evaluations LF-Antibody 4C5 can also be used to demonstrate lactoferrin molecules by conventional immunofluorescence or immunoenzyme staining techniques on cell smears, cytospin preparations or tissue sections. Acetone or paraformaldehyde are suitable fixatives for these purposes.

Commentaires:

Lactoferrin (LF) is an iron-binding protein with bactericidal and bacteriostatic activity, which is stored within the secondary granules of granulocytes. LF expression is restricted to the post-mitotic maturation compartment of the granulocytic lineage, starting from the myelocyte stage. Normal and malignant myeloblasts are LF negative. The 4C5 antibody permits the identification and enumeration of human granulocytes 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-Lactoferrin (PE) (ABIN1741598)

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).

Teng, Gladwell, Beard, Walmer, Teng, Brenner: "Lactoferrin gene expression is estrogen responsive in human and rhesus monkey endometrium." dans: Molecular human reproduction, Vol. 8, Issue 1, pp. 58-67, (2001) (PubMed).

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).

Cowland, Borregaard: "The individual regulation of granule protein mRNA levels during neutrophil maturation explains the heterogeneity of neutrophil granules." dans: Journal of leukocyte biology, Vol. 66, Issue 6, pp. 989-95, (2000) (PubMed).

Koníková, Glasová, Kusenda, Babusíková: "Intracellular markers in acute myeloid leukemia diagnosis." dans: Neoplasma, Vol. 45, Issue 5, pp. 282-91, (1999) (PubMed).

Oehler, Majdic, Pickl, Stöckl, Riedl, Drach, Rappersberger, Geissler, Knapp: "Neutrophil granulocyte-committed cells can be driven to acquire dendritic cell characteristics." dans: The Journal of experimental medicine, Vol. 187, Issue 7, pp. 1019-28, (1998) (PubMed).

Gullberg, Andersson, Garwicz, Lindmark, Olsson: "Biosynthesis, processing and sorting of neutrophil proteins: insight into neutrophil granule development." dans: European journal of haematology, Vol. 58, Issue 3, pp. 137-53, (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).

He, Furmanski: "Sequence specificity and transcriptional activation in the binding of lactoferrin to DNA." dans: Nature, Vol. 373, Issue 6516, pp. 721-4, (1995) (PubMed).

Srivastava, Rado, Bauerle, Broxmeyer: "Regulation of human bone marrow lactoferrin and myeloperoxidase gene expression by tumor necrosis factor-alpha." dans: Journal of immunology (Baltimore, Md. : 1950), Vol. 146, Issue 3, pp. 1014-9, (1991) (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).

Rado, Wei, Benz: "Isolation of lactoferrin cDNA from a human myeloid library and expression of mRNA during normal and leukemic myelopoiesis." dans: Blood, Vol. 70, Issue 4, pp. 989-93, (1987) (PubMed).

Cramer, Pryzwansky, Villeval, Testa, Breton-Gorius: "Ultrastructural localization of lactoferrin and myeloperoxidase in human neutrophils by immunogold." dans: Blood, Vol. 65, Issue 2, pp. 423-32, (1985) (PubMed).

Rado, Bollekens, St Laurent, Parker, Benz: "Lactoferrin biosynthesis during granulocytopoiesis." dans: Blood, Vol. 64, Issue 5, pp. 1103-9, (1984) (PubMed).

Détails sur Lactoferrin

Antigène: Lactoferrin (LTF) (Lactotransferrin (LTF))

Autre désignation: Lactoferrin

Pathways: Transition Metal Ion Homeostasis