CIP2A Antibody (2G10)

Western Blot: CIP2A Antibody (2G10) [NB110-59722]

Western Blot: CIP2A Antibody (2G10) [NB110-59722] - Analysis of CIP2A in NIH/3T3 cell lysate.

Immunocytochemistry/ Immunofluorescence: CIP2A Antibody (2G10) [NB110-59722]

Immunocytochemistry/Immunofluorescence: CIP2A Antibody (2G10) [NB110-59722] - CIP2A antibody was tested in U2OS cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).

Immunohistochemistry-Paraffin: CIP2A Antibody (2G10) [NB110-59722]

Immunohistochemistry-Paraffin: CIP2A Antibody (2G10) [NB110-59722] - IHC analysis of formalin fixed paraffin-embedded (FFPE) human colon cancer using CIP2A antibody at 1:200 on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Staining was performed by Histowiz.

Western Blot: CIP2A Antibody (2G10) [NB110-59722] -

Western Blot: CIP2A Antibody (2G10) [NB110-59722] - miR-375 increases p21, p53, & RB in HPV16- & 18-positive cancer. (A) Protein levels of p21, p53, & RB in SiHa cells transfected w/ miR-375 inhibitor, -mimic, or NS control measured by Western blot analysis. 25%, 50%, 100% amounts of untreated cell lysates included to calibrate the semiquantitative measurement. (B) Relative endogenous mRNA levels of p21, p53, & RB measured in SiHa cells transfected w/ miR-375 inhibitor, -mimic, or NS control using qRT-PCR. (C) One hundred thousand SiHa cells seeded on 24-well plate & the number of cells counted by trypan blue exclusion staining assay 48 h post-transfection. (D) Protein levels of p21, p53, & RB in HeLa cells transfected w/ miR-375 inhibitor, -mimic, or NS control measured by Western blot analysis. (E) Relative p21 mRNA levels measured in HeLa cells transfected w/ miR-375 inhibitor, -mimic, or NS control. (F) Trypan blue exclusion staining assay used to analyze the proliferation rate of HeLa cells 48 h post-transcfection. (G) Flow cytometry analysis demonstrates G1 arrest of SiHa cells 48 h after transfection w/ miR-375-mimic compared to miR-375 inhibitor or NS control. (H) Protein levels of CIP2A, p53, & p21 in SiHa cells transfected w/ si-CIP2A and/or si-p53 measured by Western blot analysis. (I) Protein levels of CIP2A, p53, & p21 in SiHa cells transfected w/ si-p53 and/or miR-375-mimic measured by Western blot analysis. (J) mRNA levels of p21 in SiHa cells transfected w/ si-p53 and/or miR-375-mimic measured by qRT-PCR. The concentrations of siRNA or miRNA used in panels H, I, & J 10 nM & 25 nM, respectively. Results are expressed as mean ± SD from three independent experiments. *p < 0.05, ***p < 0.001, & ****p < 0.0001. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24708873), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: CIP2A Antibody (2G10) [NB110-59722] -

Western Blot: CIP2A Antibody (2G10) [NB110-59722] - miR-375 control on CIP2A-MYC pathway also contributes to p21 elevation. (A) p21, p53, RB, CIP2A, & MYC protein levels in MCF7 cells transfected with miR-375 inhibitor, -mimic, or NS control were measured by Western blot analysis. Tubulin expression was used as internal control. 25%, 50%, 100% amounts of untreated cell lysates were included to calibrate the semiquantitative measurement. (B) Transfection with miR-375-mimic significantly upregulated p21 mRNA in MCF7. Relative endogenous p21 mRNA levels were measured in MCF7 cells transfected with miR-375 inhibitor, -mimic, or NS control for 48 h using qRT-PCR. (C) CIP2A & MYC protein levels were effectively silenced by si-CIP2A transfection with 1 & 10 nM concentrations for 48 h. Increased p21 protein levels were detected in si-CIP2A dose-dependent manner. 10 nM of si-GFP was used as a control. (D) Protein levels of CIP2A, p53, & p21 in MCF7 cells transfected with si-p53 and/or miR-375-mimic were measured by Western blot analysis. (E) mRNA levels of p21 in MCF7 cells transfected with si-p53 and/or miR-375-mimic were measured by qRT-PCR. (F) Flow cytometry analysis demonstrates G1 arrest of MCF7 cells 48 h after transfection with miR-375-mimic compared to miR-375 inhibitor or NS control. The concentrations of siRNA or miRNA used in panels D, E, & F were 10 nM & 25 nM, respectively. (G) Schematic depiction of miR-375-mediated repression of CIP2A, E6, E6AP, & E7 in HPV16-positive cells that simultaneously increases tumor suppressor p53, p21, & RB, & causes cell cycle arrest. Results are expressed as mean ± SD from three independent experiments. *p < 0.05, **p < 0.01, & ***p < 0.001. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24708873), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: CIP2A Antibody (2G10) [NB110-59722] -

Western Blot: CIP2A Antibody (2G10) [NB110-59722] - miR-375-mediated repression of HPV16, E6AP, & CIP2A activates the p53-p21 network & suppresses telomerase activity. (A) miR-375 demonstrates upregulation of p53 & p21 comparable to that of the single or combined CIP2A, E6, & E6AP knockdown. CIP2A, E6AP, p53, & p21 protein levels in SiHa cells transfected with siRNA targeting CIP2A, HPV16-E6, & E6AP (si-CIP2A, si-E6, & si-E6AP, respectively) were analyzed by Western blot. 1 nM & 10 nM siRNA concentrations & 5 nM & 50 nM for miR-375-mimic were used for transfection. si-Three is a combination of the three siRNAs indicated above. 10 nM of si-GFP was used as a control. Tubulin expression was used as internal control. (B) The increase in p21 protein levels correlate to its mRNA levels. Relative endogenous p21 mRNA levels transfected with siRNAs or miR-375 were measured using qRT-PCR. (C) miR-375 exerted a similar or stronger reduction in TERT mRNA levels when compared to E6 & E6AP knockdown in SiHa cells. (D) SiHa cells transfected with miR-375-mimic significantly reduced telomerase activity. Relative telomerase activities in SiHa cells transfected with NS control, miR-375-mimic, & miR-375 inhibitor were measured by SYBR real-time PCR TRAP assay. Heat-inactivated telomerase extracts were used to normalize this data. *p < 0.05, **p < 0.01, ***p < 0.001, & ****p < 0.0001. ns, not significant. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24708873), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: CIP2A Antibody (2G10) [NB110-59722] -

Western Blot: CIP2A Antibody (2G10) [NB110-59722] - miR-375 control on CIP2A-MYC pathway also contributes to p21 elevation. (A) p21, p53, RB, CIP2A, & MYC protein levels in MCF7 cells transfected with miR-375 inhibitor, -mimic, or NS control were measured by Western blot analysis. Tubulin expression was used as internal control. 25%, 50%, 100% amounts of untreated cell lysates were included to calibrate the semiquantitative measurement. (B) Transfection with miR-375-mimic significantly upregulated p21 mRNA in MCF7. Relative endogenous p21 mRNA levels were measured in MCF7 cells transfected with miR-375 inhibitor, -mimic, or NS control for 48 h using qRT-PCR. (C) CIP2A & MYC protein levels were effectively silenced by si-CIP2A transfection with 1 & 10 nM concentrations for 48 h. Increased p21 protein levels were detected in si-CIP2A dose-dependent manner. 10 nM of si-GFP was used as a control. (D) Protein levels of CIP2A, p53, & p21 in MCF7 cells transfected with si-p53 and/or miR-375-mimic were measured by Western blot analysis. (E) mRNA levels of p21 in MCF7 cells transfected with si-p53 and/or miR-375-mimic were measured by qRT-PCR. (F) Flow cytometry analysis demonstrates G1 arrest of MCF7 cells 48 h after transfection with miR-375-mimic compared to miR-375 inhibitor or NS control. The concentrations of siRNA or miRNA used in panels D, E, & F were 10 nM & 25 nM, respectively. (G) Schematic depiction of miR-375-mediated repression of CIP2A, E6, E6AP, & E7 in HPV16-positive cells that simultaneously increases tumor suppressor p53, p21, & RB, & causes cell cycle arrest. Results are expressed as mean ± SD from three independent experiments. *p < 0.05, **p < 0.01, & ***p < 0.001. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24708873), licensed under a CC-BY license. Not internally tested by Novus Biologicals.