Anti-KLF4 抗体

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody (AB106629)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling KLF4 with ab106629 at 2 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH 6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody (AB106629)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling KLF4 with ab106629 at 2 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH 6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody (AB106629)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling KLF4 with ab106629 at 2 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH 6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

• WB

Supplier Data

Western blot - Anti-KLF4 antibody (AB106629)

1 h incubation at RT in 5% NFDM/TBST. ab106629 detected the expression of KLF4 in MCF-7 cells, but not in Jurkat cells, which matches the RNA expression pattern in these two cell lines based on Human Protein Atlas data.

All lanes:

Western blot - Anti-KLF4 antibody (ab106629) at 1 µg/mL

Lane 1:

MCF-7 cell lysate at 15 µg

Lane 2:

Jurkat cell lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG HRP conjugate at 1/10000 dilution

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• WB

CiteAb

Western blot - Anti-KLF4 antibody (AB106629)

Western Blotting using Anti-KLF4 antibody, ab106629. Publication image from Yang, G. S. et al., 2020, Theranostics, 32685020. Legend direct from paper.

Downregulation of KLF4 by Sal-miR-1 and 3 increases NMHC IIA expression level. (A) A subset of the differentially expressed mRNAs detected in pcDNA3.1- or pcDNA3.1-KLF4-transfected VSMCs with microarray analysis was selected and summarized. (B) A schematic map of the -1843 to +42 bp region of the NMHC IIA promoter showing the positions of KLF4-binding sites. VSMCs were transfected with pcDNA3.1 or pcDNA3.1-KLF4 for 24 h, and then formaldehyde-cross-linked chromatin was extracted and immunoprecipitated with anti-KLF4 antibody. Nonimmune IgG was used as negative control for immunoprecipitation. Immunoprecipitated DNA containing KLF4 binding sites was amplified by PCR using mouse NMHC IIA promoter-specific primers. (C) VSMCs were co-transfected with KLF4 expression plasmids and NMHC IIA promoter-reporter plasmids for 24 h. Luciferase activity was measured using the dual luciferase reporter assay system. (D-G) VSMCs were transfected with pcDNA3.1-KLF4, si-KLF4 or their corresponding control for 24 h. The expression of KLF4 and NMHC IIA was analyzed by qRT-PCR (D-G) and Western blot (E-F). (H) VSMCs were transfected with Sal-miR-1+3 and/or si-OTUD7B for 24 h. The expression of OTUD7B, KLF4 and NMHC IIA was analyzed by Western blotting. All results are presented as the mean±SEM from 3 independent experiments (n=3). Student's t-test : *P<0.05, **P<0.01, ***P<0.001 versus the corresponding control.

false

• WB

CiteAb

Western blot - Anti-KLF4 antibody (AB106629)

Western Blotting using Anti-KLF4 antibody, ab106629. Publication image from Yang, G. S. et al., 2020, Theranostics, 32685020. Legend direct from paper.

OTUD7B downregulation by Sal-miR-1 and 3 attenuates KLF4 protein levels. (A) Bioinformatics analysis of the proteins interacting with OTUD7B by three protein-protein interactions databases. Images were obtained from the Cytoscape, inBio-Map and Genemania online database (Cytoscape : http : //apps.cytoscape.org/apps/agilentliteraturesearch; inBio-Map : https : //www.intomics.com/inbio/map.html#search; Genemania : http : //genemania.org). The potential proteins associated with OTUD7B were marked. (B) VSMCs were treated with 0.03 mg/mL cycloheximide (CHX), along with or without MG132 treatment. Cells were harvested at the indicated times, and KLF4 was detected by Western blotting, whereas band intensities that were measured are shown on the right. (C) VSMCs were transfected with si-Ctl or si-OTUD7B for 24 h, and the expression of OTUD7B and KLF4 was analyzed by Western blotting. (D) VSMCs were transfected with pcDNA3.1 or pcDNA3.1-OTUD7B for 24 h, and the expression of OTUD7B and KLF4 was analyzed by Western blotting. (E) VSMCs were transfected with pcDNA3.1-OTUD7B, si-OTUD7B or their corresponding control for 24 h. KLF4 expression was analyzed by qRT-PCR. (F) The Flag-tagged KLF4 expression vectors alone or together with GFP-tagged OTUD7B expression vectors were transiently transfected into HEK293A cells, after 24 h of transfection, 0.03 mg/mL CHX was added to each plate, and then incubated for 0, 2, 4, 6, and 8 h, respectively. Protein levels of KLF4 and OTUD7B were detected by Western blotting. (G) VSMCs were transfected with pcDNA3.1-OTUD7B or pcDNA3.1 for 24 h, and then anti-OTUD7B immunoprecipitates (IP) were analyzed by Western blot (WB) for OTUD7B and KLF4 co-sedimentation. IgG was used as negative control. (H) VSMCs were transfected with pcDNA3.1-OTUD7B, si-OTUD7B or their corresponding control for 24 h, and then treated with MG132 for 6 h. KLF4 was immunoprecipitated with anti-KLF4 antibody and immunoblotted with anti-ubiquitin antibody. (I) HEK293A cells were transfected for 24 h with pcDNA3.1-OTUD7B, si-OTUD7B or their corresponding control, and then treated with MG132 for 6 h. Cell lysates were incubated with anti-KLF4 antibody, and the immunoprecipitates were analyzed by immunoblotting with anti-ubiquitin antibody. All experiments were performed three independent experiments, and the results are presented as the mean ± SEM. Repeated Measures ANOVA or Student's t-test : **P<0.01, ***P<0.001 versus the corresponding control.

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