rabbit polyclonal anti rap1  (Novus Biologicals)

Journal: bioRxiv

Article Title: The ERCC6L2-MRI-KU complex coordinates NHEJ at staggered DNA double-strand breaks

doi: 10.1101/2025.11.28.691009

Figure Lengend Snippet: A. c-NHEJ at dysfunctional telomeres with and without overhang. Functional telomeres are formed by a repetitive array of TTAGGG repeats ending with a long (50-300 nt) 3’ overhang bound by the shelterin complex (TRF1, TRF2, RAP1, TIN2, TTP1, POT1). At newly replicated leading end telomeres the overhang is formed by the Exonuclease APOLLO. In the absence of TRF2, telomeres still retaining their overhang are fused by c-NHEJ (top). In the absence of APOLLO and RAP1, the blunt telomeres resulting from replication cannot be resected and are fused by c-NHEJ (bottom). B. Immunoblot for TRF2 deletion, CHK2 phosphorylation as indication of ATM activation and actin as loading control in Ercc6l2 +/+ and Ercc6l2 -/- MEFs 108 h after transduction with single guide RNA (sgRNA) targeting Trf2 (sgTrf2). C. Representative FISH of metaphase spreads of Ercc6l2 positive and negative 108 h after Trf2 deletion. Telomeres were detected with Cy3-(TTAGGG)3 (green) and DNA was stained with DAPI (magenta). Green arrows highlight chromosome-type fusions. D. Quantification of telomeres involved in chromosome fusions per metaphase after deletion of Trf2 as in (B-C). Data from 3 independent experiments, 10 metaphases per experiment (n = 30 total), with median. E. Immunoblot for Rap1 deletion, CHK2 and actin in Ercc6l2 +/+ and Ercc6l2 -/- MEFs 120 h after transduction with single guide RNA (sgRNA) targeting Rap1 (sgRAp1) and/or Apollo (sgApollo). F. Representative FISH of metaphase spreads of Ercc6l2 positive and negative MEFs 120 h after Apollo and Rap1 deletion. Telomeres were detected with Cy3-(TTAGGG)3 (green) and DNA was stained with DAPI (magenta). Green arrows highlight chromosome-type fusions. White arrows highlight chromatid-type fusions. G. Quantification of telomeres involved in chromosome fusions per metaphase after deletion of Rap1 and Apollo as in (e-f). Data from 2 (no sgRNAs) or 3 (sgRap1 + sgApollo) independent experiments, 10 metaphases per experiment with median. Ordinary one-way analysis of variance (ANOVA). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001; ns, not significant.

Article Snippet: Western blot was performed with 5% milk in PBS containing 0.1% (v/v) Tween-20 (PBS-T) using the following antibodies: β-actin (#3700; Cell Signaling), RAP1 (#5433, Cell Signalling) and TRF2 (#13136, Cell Signaling), followed by goat anti-rabbit (31460, Invitrogen) or anti-mouse (31430, Invitrogen) IgG–HRP peroxidase secondary antibody.

Techniques: Functional Assay, Western Blot, Phospho-proteomics, Activation Assay, Control, Transduction, Staining

Journal: Scientific Reports

Article Title: CD147 regulates the Rap1 signaling pathway to promote proliferation, migration, and invasion, and inhibit apoptosis in colorectal cancer cells

doi: 10.1038/s41598-025-98266-8

Figure Lengend Snippet: Information of primary antibodies used in the present study.

Article Snippet: Anti RAP1 rabbit polyclonal antibody , 14595-1-AP , ProteinTech Group, Inc. , 1:1000.

Techniques:

Journal: Scientific Reports

Article Title: CD147 regulates the Rap1 signaling pathway to promote proliferation, migration, and invasion, and inhibit apoptosis in colorectal cancer cells

doi: 10.1038/s41598-025-98266-8

Figure Lengend Snippet: Primer sequence and promoter primer sequence.

Article Snippet: Anti RAP1 rabbit polyclonal antibody , 14595-1-AP , ProteinTech Group, Inc. , 1:1000.

Techniques: Sequencing

Journal: Scientific Reports

Article Title: CD147 regulates the Rap1 signaling pathway to promote proliferation, migration, and invasion, and inhibit apoptosis in colorectal cancer cells

doi: 10.1038/s41598-025-98266-8

Figure Lengend Snippet: CD147 overexpression correlates with poor prognosis and RAP1 co-expression in colorectal cancer. ( A , B ) CD147 mRNA expression levels in tumor vs. normal tissues across multiple cancers (UCSC Xena database) and specifically in colorectal cancer (CRC) (GEPIA database). ** P < 0.01, unpaired t-test. ( C ) Kaplan-Meier survival analysis of CRC patients stratified by CD147 expression (log-rank test, P < 0.05). ( D ) Representative immunohistochemical images from The Human Protein Atlas showing CD147 expression in normal colorectal tissue (negative) and CRC tissue (cytoplasmic/membrane staining). ( E , F ) Western blot analysis of CD147 protein levels in paired CRC and adjacent normal tissues ( n = 12). β-actin served as loading control. *** P < 0.001, paired t-test. Origin blots are presented in Supplementary Fig. 1. ( G , H ) Correlation analyses between CD147 and RAP1A/B mRNA expression using GEPIA (Pearson’s correlation, P < 0.05) and TIMER2.0 (no significant correlation). ( I ) qRT-PCR validation of CD147 and Rap1 expression correlation in 12 CRC tissues (Pearson’s correlation, P < 0.01).

Article Snippet: Anti RAP1 rabbit polyclonal antibody , 14595-1-AP , ProteinTech Group, Inc. , 1:1000.

Techniques: Over Expression, Expressing, Immunohistochemical staining, Membrane, Staining, Western Blot, Control, Quantitative RT-PCR, Biomarker Discovery

Journal: Scientific Reports

Article Title: CD147 regulates the Rap1 signaling pathway to promote proliferation, migration, and invasion, and inhibit apoptosis in colorectal cancer cells

doi: 10.1038/s41598-025-98266-8

Figure Lengend Snippet: Rap1 overexpression reverses the expression of key proteins in CD147 knockdown mediated cell proliferation, apoptosis and EMT. ( A , B ) Western blot analysis of Rap1 and Rap1GAP protein levels in HCT116 and SW620 cells after CD147 knockdown (shCD147). β-actin served as a loading control. Data represent mean ± SD ( n = 3; ** P < 0.01, *** P < 0.001 vs. shNC, Student’s t-test). Original blots are presented in Supplementary Fig. 3. ( C ) qRT-PCR validation of Rap1 mRNA overexpression in CD147-knockdown cells. Expression normalized to β-actin ( n = 3; *** P < 0.001 vs. HCT116 or SW620, Student’s t-test). ( D , E ) Western blot analysis of c-Myc, Bcl-2, Bax, N-cadherin, and E-cadherin protein levels in CD147-Knockdown cells with Rap1 overexpression ( n = 3; * P < 0.05, ** P < 0.01, *** P < 0.001 vs. shNC, # P < 0.05, ## P < 0.01, ### P < 0.001 vs. shCD147, one-way ANOVA). Original blots are presented in Supplementary Fig. 4.

Article Snippet: Anti RAP1 rabbit polyclonal antibody , 14595-1-AP , ProteinTech Group, Inc. , 1:1000.

Techniques: Over Expression, Expressing, Knockdown, Western Blot, Control, Quantitative RT-PCR, Biomarker Discovery

Journal: Scientific Reports

Article Title: CD147 regulates the Rap1 signaling pathway to promote proliferation, migration, and invasion, and inhibit apoptosis in colorectal cancer cells

doi: 10.1038/s41598-025-98266-8

Figure Lengend Snippet: Rap1 overexpression rescues the regulatory effects of CD147 knockdown on proliferation and apoptosis. ( A ) CCK-8 assay showing proliferation of HCT116 ( n = 3; *** P < 0.001 vs. shNC, ### P < 0.001 vs. shCD147, one-way ANOVA). ( B , C ) Colony formation assays in HCT116 cells. Colonies were counted and normalized to shCtrl ( n = 3; ** P < 0.01, *** P < 0.001 vs. shNC, ### P < 0.001 vs. shCD147, one-way ANOVA). ( D , E ) Flow cytometry analysis of apoptosis in HCT116 cells. Percentages of apoptotic cells are shown ( n = 3; * P < 0.05 vs. shNC, # P < 0.05 vs. shCD147, one-way ANOVA). Similar results were observed in SW620 cells.

Article Snippet: Anti RAP1 rabbit polyclonal antibody , 14595-1-AP , ProteinTech Group, Inc. , 1:1000.

Techniques: Over Expression, Knockdown, CCK-8 Assay, Flow Cytometry

Journal: Scientific Reports

Article Title: CD147 regulates the Rap1 signaling pathway to promote proliferation, migration, and invasion, and inhibit apoptosis in colorectal cancer cells

doi: 10.1038/s41598-025-98266-8

Figure Lengend Snippet: Rap1 restores CD147-mediated migration and invasion in colorectal cancer cells. ( A , C ) Scratch wound healing assay in HCT116 cells. Healing rates were quantified at 24 h and 48 h ( n = 3; *** P < 0.001 vs. shNC, ## P < 0.01, ### P < 0.001 vs. shCD147, one-way ANOVA). ( B , D ) Transwell migration and Matrigel invasion assays. Migrated/invaded cells were counted and normalized to shCtrl ( n = 3; *** P < 0.001 vs. shNC, ### P < 0.001 vs. shCD147, one-way ANOVA). SW620 cells showed consistent trends. ( E ) Mechanistic diagram of CD147 promoting tumor progression through Rap1/Rap1GAP signaling in colorectal cancer cells.

Article Snippet: Anti RAP1 rabbit polyclonal antibody , 14595-1-AP , ProteinTech Group, Inc. , 1:1000.

Techniques: Migration, Wound Healing Assay

Journal: Epigenetics & Chromatin

Article Title: Ubiquitin C-terminal hydrolase isozyme L1 is associated with shelterin complex at interstitial telomeric sites

doi: 10.1186/s13072-017-0160-2

Figure Lengend Snippet: UCHL1 interacts with the shelterin complex and the nuclear scaffold. a DU 145 or HEK293T cellular lysates in a low stringency buffer were incubated with anti-TRF2 antibodies or control IgG. The immunoprecipitate (IP), and equal volumes of lysate (Input) and immunodepleted (ID) fractions were analyzed by immunoblotting with the indicated antibodies. b In left panel, DU 145 cellular lysate in a low stringency buffer was incubated with anti-p53 antibodies or control IgG. In right panel, cell lysate in a high stringency buffer of DU 145 cells treated with the cross-linker DSP was incubated with anti-UCHL1 antibodies or control IgG. The immunoblot analysis was done as in ( a ). c In situ PLA images of the interaction between TRF2 and RAP1 (positive control), UCHL1 and RAP1 or UCHL1 and TRF2 in HEK293T cells. PLA signals appear as discrete red dots and nuclei are visualized by DAPI (blue). A total of 30 nuclei per group were quantified. The average number of PLA foci per nucleus was graphed with error bars representing standard errors of the means. Single primary antibodies, isotype control and PLA probes only were used as negative controls as indicated. **** p < 0.0001 (determined by one-way ANOVA)

Article Snippet: Antibodies used were anti-UCHL1 rabbit monoclonal (Abcam), anti-TRF2 mouse monoclonal (Novus Biologicals), anti-RAP1 (TERF2IP) mouse monoclonal (Abcam), anti-RAP1 (TERF2IP) rabbit polyclonal (Bethyl Laboratories), rabbit IgG and mouse IgG (Sigma-Aldrich).

Techniques: Incubation, Control, Western Blot, In Situ, Positive Control

Journal: Epigenetics & Chromatin

Article Title: Ubiquitin C-terminal hydrolase isozyme L1 is associated with shelterin complex at interstitial telomeric sites

doi: 10.1186/s13072-017-0160-2

Figure Lengend Snippet: UCHL1 and RAP1 are associated with the nuclear scaffold. a Proteins (5 µg) from each fraction [total cellular protein (TC), total nuclear protein (TN), ammonium sulfate fraction (AS) and nuclear scaffold protein (NS)] were resolved on a SDS 15% polyacrylamide gel and stained with Coomassie Blue. The DU 145 cells were treated or not with protein–protein cross-linker DSP. b Immunoblot analysis using the indicated antibodies on TC, TN, AS and NS proteins from DU 145 cells treated or not with protein–protein cross-linker DSP. PC-3 cells were not treated with DSP. c Immunoblot analysis of UCHL1 and RAP1 in nuclear scaffold protein complexes from DSP-treated DU 145 cells. The nuclear scaffold was solubilized in SDS sample buffer in both reducing (R) and non-reducing (NR) conditions and analyzed by SDS-PAGE on a 10% gels and immunoblotting

Article Snippet: Antibodies used were anti-UCHL1 rabbit monoclonal (Abcam), anti-TRF2 mouse monoclonal (Novus Biologicals), anti-RAP1 (TERF2IP) mouse monoclonal (Abcam), anti-RAP1 (TERF2IP) rabbit polyclonal (Bethyl Laboratories), rabbit IgG and mouse IgG (Sigma-Aldrich).

Techniques: Staining, Western Blot, SDS Page

Journal: Epigenetics & Chromatin

Article Title: Ubiquitin C-terminal hydrolase isozyme L1 is associated with shelterin complex at interstitial telomeric sites

doi: 10.1186/s13072-017-0160-2

Figure Lengend Snippet: UCHL1 knockdown and shelterin protein levels. HEK293T cells were transfected with GAPDH siRNA (50 nM), scrambled siRNA control (50 nM) and two UCHL1 siRNA with the final concentrations of 25 and 50 nM. Whole protein lysates were extracted from control and siRNA-transfected HEK293T cells 72 h posttransfection, and immunoblot analysis was performed with antibodies against UCHL1, RAP1, TRF2 and GAPDH. Beta actin was used as an internal control

Article Snippet: Antibodies used were anti-UCHL1 rabbit monoclonal (Abcam), anti-TRF2 mouse monoclonal (Novus Biologicals), anti-RAP1 (TERF2IP) mouse monoclonal (Abcam), anti-RAP1 (TERF2IP) rabbit polyclonal (Bethyl Laboratories), rabbit IgG and mouse IgG (Sigma-Aldrich).

Techniques: Knockdown, Transfection, Control, Western Blot