Recombinant Anti-KLF5 antibody [23HCLC] (ab277773)

For immunofluorescence analysis, A549 cells were fixed and permeabilized for detection of endogenous KLF5 using Anti- KLF5 Recombinant Rabbit Multiclonal Antibody (ab277773, 2 μ/mL) and labeled with Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor^® 488 conjugate (1/2000). Panel a) shows representative cells that were stained for detection and localization of KLF5 protein (green), Panel b) is stained for nuclei (blue) with DAPI. Panel c) represents cytoskeletal F-actin staining using Alexa Fluor^® 555 Rhodamine Phalloidin (1/300). Panel d) is a composite image of Panels a, b and c demonstrating predominant nuclear localization of KLF5. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Enrichment of endogenous KLF5 protein at specific gene loci using Anti- KLF5 Recombinant Rabbit Multiclonal Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-KLF5 Recombinant Rabbit Multiclonal Antibody (ab277773, 5 μg) on sheared chromatin from 2 million A549 cells. Normal Rabbit IgG (1 μg) was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system with optimized PCR primer pairs for the promoters of the active SOX4, KLF4 region used as positive control target gene, and the region of the inactive KLF4 UPSTRAEM, SAT2 satellite repeat, used as negative control target gene. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.