Mouse anti-Human Lactoferrin Antibody | anti-LTF antibody
Mouse anti Lactoferrin
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 ul of leukocytes containing 10^7
cells/ml are stained with 20 ul 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.
- In combination with our Permeabilization Kit FIX&PERM intracellular Lactoferrin can be easily stained in cell suspensions.
- For each sample to be analyzed add 50 ul of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube
- Add 100 ul of Reagent A (Fixation Medium, stored and used at room temperature)
- Incubate for 15 minutes at room temperature
- Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g
- Remove supernatant and add to cell pellet 100 ul Reagent B (Permeabilization Medium) and 20 ul 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 degree 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.
The 4C5 antibody permits the identification and enumeration of human granulocytes using flow cytometry. Results must be put within the context of other tests as well as the clinical history of the patient by a certified professional before final interpretation. Analysis performed with this antibody should be paralleled by positive and negative controls.
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NCBI and Uniprot Product Information
NCBI Description
This gene is a member of the transferrin family of genes and its protein product is found in the secondary granules of neutrophils. The protein is a major iron-binding protein in milk and body secretions with an antimicrobial activity, making it an important component of the non-specific immune system. The protein demonstrates a broad spectrum of properties, including regulation of iron homeostasis, host defense against a broad range of microbial infections, anti-inflammatory activity, regulation of cellular growth and differentiation and protection against cancer development and metastasis. Antimicrobial, antiviral, antifungal and antiparasitic activity has been found for this protein and its peptides. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Sep 2014]
Uniprot Description
LTF: Transferrins are iron binding transport proteins which can bind two Fe(3+) ions in association with the binding of an anion, usually bicarbonate. Belongs to the transferrin family. 2 isoforms of the human protein are produced by alternative promoter.
Protein type: Secreted, signal peptide; Protease; Secreted; EC 3.4.21.-
Chromosomal Location of Human Ortholog: 3p21.31
Cellular Component: extracellular space; specific granule; cytoplasm; extracellular region; nucleus; secretory granule
Molecular Function: heparin binding; protein serine/threonine kinase activator activity; protein binding; ferric iron binding; DNA binding; iron ion binding; serine-type endopeptidase activity
Biological Process: interaction with host; negative regulation of lipopolysaccharide-mediated signaling pathway; ossification; positive regulation of I-kappaB kinase/NF-kappaB cascade; transcription, DNA-dependent; positive regulation of toll-like receptor 4 signaling pathway; antifungal humoral response; proteolysis; iron ion transport; humoral immune response; regulation of cytokine production; activation of NF-kappaB transcription factor; retinal homeostasis; positive regulation of osteoblast differentiation; innate immune response in mucosa; negative regulation of viral genome replication; response to host immune response; positive regulation of osteoblast proliferation; antibacterial humoral response; regulation of tumor necrosis factor production; negative regulation of ATPase activity; iron assimilation by chelation and transport; negative regulation of viral reproduction; negative regulation of apoptosis
Research Articles on LTF
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Product Notes
The LTF ltf (Catalog #AAA570346) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Mouse anti Lactoferrin reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's Lactoferrin can be used in a range of immunoassay formats including, but not limited to, Flow Cytometry (FC/FACS), Indirect Immunofluorescence (IF). Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM intracellular Lactoferrin can be easily stained in cell suspensions. - For each sample to be analyzed add 50 ul of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube - Add 100 ul of Reagent A (Fixation Medium, stored and used at room temperature) - Incubate for 15 minutes at room temperature - Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g - Remove supernatant and add to cell pellet 100 ul Reagent B (Permeabilization Medium) and 20 ul 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 degree 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. Researchers should empirically determine the suitability of the LTF ltf for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Lactoferrin, Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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