Anti-TPX2 Antibody Picoband®

Western blot analysis of TPX2 using anti-TPX2 antibody (A01610-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human COLO-320 whole cell lysates,
Lane 3: human U-87MG whole cell lysates,
Lane 4: human SGC-7901 whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: mouse HEPA1-6 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-TPX2 antigen affinity purified polyclonal antibody (Catalog # A01610-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for TPX2 at approximately 86KD, 100KD. The expected band size for TPX2 is at 86KD.

HMGB3 could inhibit cell survival via TPX2 in NB cells. (A) Nine genes significantly co-expressed with HMGB3 in four datasets of NB. (B) Functional enrichment analysis of nine genes. (C) The mRNA expression of nine genes was detected by qRT-PCR in SK-N-SH cells after HMGB3 knockdown. (D) Pearson correlation of HMGB3 and TPX2 in four NB datasets. (E) Protein expression of TPX2 and HMGB3 was detected by western blotting after HMGB3 knockdown. (F) TPX2 reduction was time-dependent with HMGB3 knockdown in SK-N-SH and SK-N-AS cells. * p < 0.05, ** p < 0.01, and *** p < 0.001.
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TPX2 overexpression can reverse the inhibition of cell proliferation caused by HMGB3 knockdown in SK-N-SH cells. (A) Protein expression was detected in sh-HMGB3 SK-N-SH cells after TPX2 over-expression by western blotting. (B–D) Colon formation, Transwell assay, and immunocytochemistry assay for sh-HMGB3 SK-N-SH cells after TPX2 over-expression. Scale bars, 50 μm * p < 0.05, ** p < 0.01, and *** p < 0.001.
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High expression of HMGB3 and TPX2 correlated to inferior NB prognosis in multiple datasets. (A–C) , Overall survival (OS) of NB patients in three independent datasets. (D, E) Progression-free survival (PFS) and Event-free survival (EFS) in NB patients.
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HMGB3 and TPX2 are closely associated with the survival of patients with NB in GSE49710. (A) HMGB3 and TPX2 can be independent prognostic factors for overall survival. (B–D) Nomogram, random forest, and ROC curves of HMGB3, TPX2 and clinicopathological characteristics for NB patients.
Index in PubMed under a CC BY license. PMID: 34988076

CircPOSTN regulated TPX2 expression via sponging miR-361-5p in glioma cells. a The binding sequences between miR-361-5p and TPX2, and matched mutant sites were shown. b , c Luciferase activity was determined by dual-luciferase reporter assay in LN229 and U251 cells following co-transfection with TPX2 3′UTR-WT or TPX2 3′UTR-MUT and miR-361-5p or miR-NC according to the design. d – g The mRNA and protein expression levels of TPX2 were estimated using RT-qPCR and western blot assays in glioma tissues and cells, along with controls. h , i The correlation analysis between TPX2 and miR-361-5p or circPOSTN was conducted by Pearson’s correlation analysis. j , k The RT-qPCR and western blot assays were recruited to evaluate the expression levels of TPX2 level in LN229 and U251 cells transfected with miR-NC, miR-361-5p, miR-361-5p + pcDNA-NC or miR-361-5p + pcDNA-circPOSTN. * P < 0.05
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TPX2 regulated proliferation, apoptosis, and aerobic glycolysis in glioma cells. a – l LN229 and U251 cells were introduced with si-NC or si-TPX2. a The transfection efficiency of si-TPX2 was checked with RT-qPCR assay in LN229 and U251 cells. b , c The cell viability of LN229 and U251 cells was determined with MTT assay. d The apoptosis rate of transfected LN229 and U251 cells was represented by flow cytometry assay. e The western blot assay was used to assay the expression levels of Bcl-2 and Bax in LN229 and U251 cells. f The activity of caspase-3 was detected with a caspase-3 assay kit. g – i The glucose, lactate, and ATP production levels were shown. j The protein expression levels of HK2 and LDHA were estimated by western blot assay in LN229 and U251 cells. k , l LDHA enzyme activity and ROS content were evaluated in LN229 and U251 cells post-transfection. * P < 0.05
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Silencing of circPOSTN repressed glioma tumor growth in vivo. a , b Tumor volume and weight were presented. c , d The expression levels of circPOSTN and miR-361-5p in dissected tumor tissues were estimated with RT-qPCR assay. e Western blot assay was executed to test protein level of TPX2 in dissected tumor tissues. f The diagrammatic representation of our conclusion was shown. * P < 0.05
Index in PubMed under a CC BY license. PMID: 32774168