Immunofluorescence analysis of KLF4 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with KLF4 Rabbit Polyclonal Antibody (PA1095) at 1ug/ml in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant N-terminal fragment (amino acids 1-118) of human KLF4|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||1:500-1:2000|
* 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.
The predicted molecular weight of KLF4 is ~55kD. The migration of this protein by SDS-PAGE was observed slightly higher (~60kD) with both PA1-095 and a benchmark antibody.
PA1-095 detects a predominant band at ~60kD, and a nonspecific band of unknown origin at ~80kD in both whole cell lysates and nuclear extracts.
For best results, StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) is recommended.
KLF4 (Kruppel-like factor 4 (gut)) is a transcription factor that can act both as an activator and as a repressor. KLF4 binds to the 5'-CACCC-3' core sequence, and binds to the promoter region of its own gene and it can activate its own transcription. It regulates the expression of key transcription factors during embryonic development. KLF4 plays an important role in maintaining embryonic stem cells, and in preventing their differentiation. It is required for establishing the barrier function of the skin and for postnatal maturation and maintenance of the ocular surface. It is also involved in the differentiation of epithelial cells and may also function in skeletal and kidney development. Diseases associated with KLF4 include skin sequamous cell carcinoma and familial adenomatous polyposis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.