Immunofluorescence analysis of GCSF (BVD13 3A5) was performed using 70% confluent log phase K562 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 G-CSF BVD13-3A5) Rat Monoclonal Antibody (Product # AHC2034) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A11006) 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 (Product # 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 no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rat / IgG1|
|Immunogen||Recombinant human G-CSF expressed in E. coli.|
|Storage buffer||PBS, pH 7.2|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||Assay Dependent|
* 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.
Granulocyte colony-stimulating factors promotes survival, proliferation and differentiation of bone marrow precursor cells. GCSF is a lineage-restricted hematopoietic growth factor, stimulating final mitotic divisions and terminal cellular maturation of partially differentiated hematopoietic progenitors.
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