Mouse anti Lactoferrin, conjugated to PE

Nordic MuBio
Product Code: GM-4113
Product Group: Primary Antibodies
Supplier: Nordic MuBio
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GM-41132ml (100 Tests)£372.00
Quantity:
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Overview

Host Type: Mouse
Antibody Isotype: IgG1
Antibody Clonality: Monoclonal
Antibody Clone: 4C5
Regulatory Status: (Not IVD)
Target Species: Human
Applications:
  • Flow Cytometry
  • Immunofluorescence (IF)
Storage:
Nordic-MUbio monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody. For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8°C (DO NOT FREEZE!) and protec
Keywords:
Nordic MuBio

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Further Information

Applications Description:
Permeabilization and Staining Procedure
- In combination with our Permeabilization Kit FIX&PERM? (Cat. NoGAS-002) intracellular Lactoferrin can be easily stained in cell suspensions.
- For each sample to be analyzed add 50 ?l of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube
- Add 100 ?l 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 ?l Reagent B (Permeabilization Medium) and 20 ?l 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.
Background:
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. If unexpected results are obtained which cannot be attributed to differences in laboratory procedures, please contact us.
Caution:
For professional users only.
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.
Formulation:
Purified by Chromatography, Storage buffer: PBS pH 7.2, 1% BSA, 0.05% NaN3
Label:
PE
Product:
2 ml of PE-conjugated anti Lactoferrin (clone 4C5) in PBS pH 7.2, 1% BSA, and 0.05% NaN3, approximately 100 tests.
Product Form:
PE
Specificity:
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 ?l of leukocytes containing 10^7
cells/ml are stained with 20 ?l 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.

References

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