tcp11 antibody  (Proteintech)

Journal: BMC cancer

Article Title: Increased TCP11 gene expression can inhibit the proliferation, migration and promote apoptosis of cervical cancer cells.

doi: 10.1186/s12885-023-11129-1

Figure Lengend Snippet: Fig. 1 TCP11 gene is highly expressed in cervical cancer tissues and cells, and is closely associated with patients’ survival rate. A: The mRNA expression of TCP11 in normal cervical tissues and cervical cancer tissues were analyzed by GEPIA database. B: Immunohistochemical results of TCP11 protein expres sion in normal cervical tissues (n = 31) and cervical cancer tissues (n = 35) microarray. “-” indicates that the expression of TCP11 is negative; “+” indicates that the expression of TCP11 is weakly positive; “++” indicates that the expression of TCP11 is medium positive. Magnification: ×100 (top) and ×400 (bottom). C: Western blot was used to detect the expression of TCP11 protein in immortalized epithelial cells HaCaT and three cervical cancer cells. Full-length blots/ gels are presented in Supplementary Figure S1. D: qRT-PCR was used to detect the expression of TCP11 mRNA in immortalized epithelial cells HaCaT and three cervical cancer cells. E: GEPIA database was used to analyze the relationship between the expression of TCP11 and survival rate of cervical cancer patients. Data are presented as mean ± SD of at least 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Article Snippet: After incubation in the dark in 3% H2O2, they were incubated with TCP11 antibody (Proteintech, China, 1:200) overnight.

Techniques: Expressing, Immunohistochemical staining, Microarray, Western Blot, Quantitative RT-PCR

Journal: BMC cancer

Article Title: Increased TCP11 gene expression can inhibit the proliferation, migration and promote apoptosis of cervical cancer cells.

doi: 10.1186/s12885-023-11129-1

Figure Lengend Snippet: Fig. 2 TCP11 overexpression inhibits the proliferation of HeLa and SiHa cells. A: Western blot was used to detect the overexpression of TCP11 protein in HeLa and SiHa cells after lentivirus infection. Full-length blots/gels are presented in Supplementary Figure S2. B: qRT-PCR was used to detect the overex pression of TCP11 mRNA in HeLa and SiHa cells after lentivirus infection. C and D: The cell viability of HeLa and SiHa cells was detected by MTT and colony formation assays. E: The protein expression of Ki67 in HeLa and SiHa cells was detected by cellular immunohistochemistry. Magnification: ×100. The results were analyzed by Image Pro Plus software, and the mean density was calculated (mean density = IOD/area). F: Ki67 mRNA expression in HeLa and SiHa cells were detected by qRT-PCR. Data are presented as mean ± SD of at least 3 independent experiments. *P < 0.05, **P < 0.01, ****P < 0.0001

Article Snippet: After incubation in the dark in 3% H2O2, they were incubated with TCP11 antibody (Proteintech, China, 1:200) overnight.

Techniques: Over Expression, Western Blot, Infection, Quantitative RT-PCR, Expressing, Immunohistochemistry, Software

Journal: BMC cancer

Article Title: Increased TCP11 gene expression can inhibit the proliferation, migration and promote apoptosis of cervical cancer cells.

doi: 10.1186/s12885-023-11129-1

Figure Lengend Snippet: Fig. 3 TCP11 overexpression blocks cell cycle progression in HeLa and SiHa cells. A and B: Cell cycle distribution of HeLa and SiHa cells was determined by flow cytometry. C: Western blot was used to detect the protein expression of CDK1 and Cyclin B1 in HeLa and SiHa cells infected with overexpressing TCP11 lentivirus. Full-length blots/gels are presented in Supplementary Figure S3. D: qRT-PCR was used to detect the mRNA expression of CDK1 and Cyclin B1 in HeLa and SiHa cells infected with overexpressing TCP11 lentivirus. Data are presented as mean ± SD of at least 3 independent experiments. *P < 0.05, **P < 0.01, ****P < 0.0001

Article Snippet: After incubation in the dark in 3% H2O2, they were incubated with TCP11 antibody (Proteintech, China, 1:200) overnight.

Techniques: Over Expression, Flow Cytometry, Western Blot, Expressing, Infection, Quantitative RT-PCR

Journal: BMC cancer

Article Title: Increased TCP11 gene expression can inhibit the proliferation, migration and promote apoptosis of cervical cancer cells.

doi: 10.1186/s12885-023-11129-1

Figure Lengend Snippet: Fig. 4 TCP11 overexpression induces the apoptosis of HeLa and SiHa cells. A and B: Apoptosis of HeLa and SiHa cells were detected by flow cytometry. C: Western blot was used to detect the protein expression of caspase-3, cleaved-caspase-3 and cleaved-PARP in HeLa and SiHa cells infected with over expressing TCP11 lentivirus. Full-length blots/gels are presented in Supplementary Figure S4. Data are presented as mean ± SD of at least 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Article Snippet: After incubation in the dark in 3% H2O2, they were incubated with TCP11 antibody (Proteintech, China, 1:200) overnight.

Techniques: Over Expression, Flow Cytometry, Western Blot, Expressing, Infection

Journal: BMC cancer

Article Title: Increased TCP11 gene expression can inhibit the proliferation, migration and promote apoptosis of cervical cancer cells.

doi: 10.1186/s12885-023-11129-1

Figure Lengend Snippet: Fig. 5 TCP11 overexpression inhibits cervical cancer cell migration by inhibiting EMT of HeLa and SiHa cells. A and B: Cell scratch assay was used to detect the migration ability of HeLa and SiHa cells. C and D: Transwell migration assay was used to detect the migration ability of HeLa and SiHa cells. E: Western blot was used to detect the protein expression of EMT-related molecules ZO-1, E-cadherin, Claudin-1, Snail, Vimentin and β-catenin in HeLa and SiHa cells infected with overexpressing TCP11 lentivirus. Full-length blots/gels are presented in Supplementary Figure S5. F: qRT-PCR was used to detect the mRNA expression of EMT-related molecules ZO-1 and E-cadherin in HeLa and SiHa cells infected with overexpressing TCP11 lentivirus. Data are presented as mean ± SD of at least 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Article Snippet: After incubation in the dark in 3% H2O2, they were incubated with TCP11 antibody (Proteintech, China, 1:200) overnight.

Techniques: Over Expression, Migration, Wound Healing Assay, Transwell Migration Assay, Western Blot, Expressing, Infection, Quantitative RT-PCR

Journal: BMC cancer

Article Title: Increased TCP11 gene expression can inhibit the proliferation, migration and promote apoptosis of cervical cancer cells.

doi: 10.1186/s12885-023-11129-1

Figure Lengend Snippet: Fig. 6 TCP11 knockdown promotes the proliferation of HeLa and SiHa cells and the migration of HeLa cells. A: Western blot was used to detect the knock down effect of three designed and constructed siRNA fragments on TCP11 in HeLa and SiHa cells. Full-length blots/gels are presented in Supplementary Figure S6. B: qRT-PCR was used to detect the knockdown effect of three siRNA fragments on TCP11 in HeLa and SiHa cells. C: MTT assay was used to detect the effect of TCP11 knockdown on the proliferation of HeLa and SiHa cells. D: Colony formation assay was used to detect the effect of TCP11 knockdown on the proliferation of HeLa cells. E: Transwell migration assay was used to detect the effect of TCP11 knockdown on the migration of HeLa cells. Data are presented as mean ± SD of at least 3 independent experiments. *P < 0.05, **P < 0.01, ****P < 0.0001

Article Snippet: After incubation in the dark in 3% H2O2, they were incubated with TCP11 antibody (Proteintech, China, 1:200) overnight.

Techniques: Knockdown, Migration, Western Blot, Construct, Quantitative RT-PCR, MTT Assay, Colony Assay, Transwell Migration Assay

Journal: BMC Cancer

Article Title: Increased TCP11 gene expression can inhibit the proliferation, migration and promote apoptosis of cervical cancer cells

doi: 10.1186/s12885-023-11129-1

Figure Lengend Snippet: TCP11 overexpression induces the apoptosis of HeLa and SiHa cells. A and B : Apoptosis of HeLa and SiHa cells were detected by flow cytometry. C : Western blot was used to detect the protein expression of caspase-3, cleaved-caspase-3 and cleaved-PARP in HeLa and SiHa cells infected with overexpressing TCP11 lentivirus. Full-length blots/gels are presented in Supplementary Figure . Data are presented as mean ± SD of at least 3 independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Anti-TCP11 (14606-1-AP) and anti-caspase-3 (8193T) were purchased from Proteintech (China).

Techniques: Over Expression, Flow Cytometry, Western Blot, Expressing, Infection

Journal: Biology of Reproduction

Article Title: Mouse t-complex protein 11 is important for progressive motility in sperm ^†

doi: 10.1093/biolre/ioz226

Figure Lengend Snippet: Tcp11 is evolutionary conserved and expressed in testis. (A) Multiple sequence alignment of human, mouse, and rat TCP11. Darker color represents greater conservation. (B) Two verified variants of mouse TCP11 are annotated, one 567 amino acids long and a shorter version 488 amino acids long. Two additional TCP11 variants are predicted that are shorter. Most of TCP11 is composed of the TCP11 domain (black), which is specific to TCP11 homologs and is uncharacterized. Grey highlights the region specific to variant 1. Red highlights the region used to generate an antibody. (C) RT-PCR from various mouse tissues detects expression of Tcp11 in testis beginning at post-natal day 15. Hprt (hypoxanthine-guanine phosphoribosyltransferase) was used as a control.

Article Snippet: The Novus anti-TCP11 antibody (NBP1-57698) was used at 1:500 dilution for Western blot analysis [ ].

Techniques: Sequencing, Variant Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Control

Journal: Biology of Reproduction

Article Title: Mouse t-complex protein 11 is important for progressive motility in sperm ^†

doi: 10.1093/biolre/ioz226

Figure Lengend Snippet: Tcp11 -null males are subfertile. (A) Schematic of mouse Tcp11 , the targeted Tcp11 tm1a(WTSI) floxed allele ( Tcp11 fl ), and the post-cre null allele Tcp11 tm1b ( Tcp11 − ). (B) Genotyping PCR distinguishing wild-type, heterozygous Tcp11 +/− , and Tcp11 knockout ( Tcp11 −/− ) mice. (C) Western blot analysis with an antibody raised against a region near the N-terminus, detects both the longer and shorter variants of TCP11 in wild-type testis lysate but not in epididymal lysates. IZUMO1 (a component of the acrosome) confirms the presence of spermatozoa in both the testis and epididymal lysate. (D) Mating tests show Tcp11 −/− sire fewer pups over a three-month mating period than controls when paired with wild-type females ( n = 5 males).

Article Snippet: The Novus anti-TCP11 antibody (NBP1-57698) was used at 1:500 dilution for Western blot analysis [ ].

Techniques: Knock-Out, Western Blot

Journal: Biology of Reproduction

Article Title: Mouse t-complex protein 11 is important for progressive motility in sperm ^†

doi: 10.1093/biolre/ioz226

Figure Lengend Snippet: Spermatogenesis appears normal in the absence of Tcp11 . (A) Gross morphology of testis from control (het) and Tcp11 KO males. (B) Average testis weight from control and Tcp11 KO adult males. (C and D) PAS stained testis cross sections from control and Tcp11 KO mice, respectively. Scale bars = 50 μm. (E and F) PAS stained epididymal sections from control and Tcp11 KO mice, respectively. Scale bars = 50 μm.

Article Snippet: The Novus anti-TCP11 antibody (NBP1-57698) was used at 1:500 dilution for Western blot analysis [ ].

Techniques: Control, Staining

Journal: Biology of Reproduction

Article Title: Mouse t-complex protein 11 is important for progressive motility in sperm ^†

doi: 10.1093/biolre/ioz226

Figure Lengend Snippet: Tcp11 KO sperm exhibit aberrant morphology and reduced motility. (A and B) Phase contrast microscopy images of sperm isolated from the cauda epididymis from wild type and Tcp11 KO. (C) Percent of sperm isolated from the cauda epididymis displaying abnormal morphology between wild type and Tcp11 KO. (D) Percent of sperm that are motile and progressively motile in wild type and Tcp11 KO determined by CASA analysis after incubation in TYH medium for 10 and 120 min. (E) Motility parameters of sperm from wild type and Tcp11 KO determined by CASA analysis after incubation in TYH medium for 10 and 120 min. ( * 0.05 > P > 0.001; ** P < 0.001).

Article Snippet: The Novus anti-TCP11 antibody (NBP1-57698) was used at 1:500 dilution for Western blot analysis [ ].

Techniques: Microscopy, Isolation, Incubation

Journal: Biology of Reproduction

Article Title: Mouse t-complex protein 11 is important for progressive motility in sperm ^†

doi: 10.1093/biolre/ioz226

Figure Lengend Snippet: Tcp11 KO sperm can fertilize oocytes without the zona pellucida. Fertilization rates using wild-type or Tcp11 KO sperm obtained from the cauda epididymis. CI, cumulus intact; CF, cumulus free; ZF, zona free ( ** P < 0.001).

Article Snippet: The Novus anti-TCP11 antibody (NBP1-57698) was used at 1:500 dilution for Western blot analysis [ ].

Techniques:

Journal: Biology of Reproduction

Article Title: Mouse t-complex protein 11 is important for progressive motility in sperm ^†

doi: 10.1093/biolre/ioz226

Figure Lengend Snippet: TCP11 localizes to the cytosol in late spermiogenesis. (A and B) Anti-TCP11 antibody (green) displays cytoplasmic localization in testis cross sections from wild-type mice. In Tcp11 KO testis cross sections, anti-TCP11 antibody staining (green) is punctate and present in all cell types. Anti-acetylated-TUBULIN (magenta) localizes to the flagellum of spermatids in both wild-type and KO cross sections. White = Hoechst 33342. Scale bars = 50 μm. (C and D) Anti-TCP11 antibody (green) displays cytoplasmic localization in testis cross sections from wild-type mice. In Tcp11 KO testis cross sections, anti-TCP11 antibody staining (green) is diffuse in the testis and brightly stains the interstitial cells. Anti-IZUMO1 (magenta) localizes to the developing acrosome in round and elongated spermatids in both wild-type and KO cross sections. White = Hoechst 33342. Scale bars = 50 μm.

Article Snippet: The Novus anti-TCP11 antibody (NBP1-57698) was used at 1:500 dilution for Western blot analysis [ ].

Techniques: Staining

Journal: Biology of Reproduction

Article Title: Mouse t-complex protein 11 is important for progressive motility in sperm ^†

doi: 10.1093/biolre/ioz226

Figure Lengend Snippet: TCP11 appears to not be present in mature sperm. Sperm isolated from the cauda epididymis were used for protein extraction into a Triton X-100- (membrane bound and cytoplasmic soluble), SDS- (axonemal components), and SDS-insoluble (fibrous sheath and outer dense fibers) pool. Western blot analysis against BASIGIN, acetylate-Tubulin, and AKAP4 demonstrates successful extraction of these three pools, respectively. Anti-TCP11 antibody only detects bands in the wild-type testis lysate.

Article Snippet: The Novus anti-TCP11 antibody (NBP1-57698) was used at 1:500 dilution for Western blot analysis [ ].

Techniques: Isolation, Protein Extraction, Membrane, Western Blot, Extraction

Journal: Biology of Reproduction

Article Title: Mouse t-complex protein 11 is important for progressive motility in sperm ^†

doi: 10.1093/biolre/ioz226

Figure Lengend Snippet: Tcp11 KO sperm have decreased phosphorylation of PKA substrates. Western blot analysis using anti-PKA substrate phosphorylation antibody shows decreased signal in sperm proteins isolated from three Tcp11 KOs. Sperm were incubated for 10 min in TYH medium prior to protein extraction.

Article Snippet: The Novus anti-TCP11 antibody (NBP1-57698) was used at 1:500 dilution for Western blot analysis [ ].

Techniques: Phospho-proteomics, Western Blot, Isolation, Incubation, Protein Extraction