Immunofluorescence analysis of Lactoferrin was performed using 70% confluent log phase HEL 92.1.7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Lactoferrin (1B8) Mouse Monoclonal Antibody (LFMA0109) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the control without primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant human lactoferrin expressed in yeast pichia pastoris|
|Purification||Ammonium sulfate precipitation|
|Storage buffer||HEPES with 0.15M NaCl, 0.01% BSA, 50% glycerol|
|Contains||0.03% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||Assay dependent|
|Immunoprecipitation (IP)||1-2 µl|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
A suggested positive control for this product is human Lactoferrin purified from first breast milk.
Lactoferrin (Lf), one of transferrin family, is found in external fluids, including milk and mucosal secretions, and prominent components of the secondary granules of neutrophils. Lf consists of a single polypeptide chain (approximately 80 kDa) folded into two structurally homologous lobes, each of which can reversibly bind one ferric ion (Fe3+). Lf plays a central role in iron metabolism and host defense system against microbial infection.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.