anti aspm rabbit polyclonal antibody  (Bethyl)

Journal: Future science OA

Article Title: Adverse predictive value of ASPM on lung adenocarcinoma overall survival depended on chemotherapy status.

doi: 10.1080/20565623.2025.2489328

Figure Lengend Snippet: Figure 1. (A) TCGA TARGET GTEx pan-cancer dataset analysis showed the relationship between ASPM gene expression and patient prognosis in each tumor. (B) In TCGA dataset of LUAD, high ASPM expression was related to worse overall survival (Log-rank test p = 2.4e-6). (C) In proteomics dataset of LUAD, the difference of prognosis was not significant between ASPM high or low expression groups (Log-rank test p = 0.26).

Article Snippet: Immunohistochemistry and special staining the Luad tissues (n = 160) were obtained from surgically resected specimens and made into tissue microarray. the sections were de-paraffinized and hydrated, and the endogenous peroxidase was blocked. antigen retrieval was performed using the dako target Retrieval Solution, High pH (dako ominis, agilent technologies, Santa Clara, Ca, uSa), in a ptLink set at 98 °C for 25 min. the tissue sections were incubated with an anti-aSpm rabbit polyclonal antibody (1:200, a02584-1, Boster, China) 1h at 37 °C. detection of immunostaining was achieved by an enzyme-conjugated polymer complex (K8002, dako, denmark) adapted for autostainers from daKo (dako autostainer, agilent technologies). the staining intensity in selected areas was scored as follows: 0, no staining; 1, weak staining; 2, moderate staining; 3, strong staining. the stained area was scored as follows: 0, no staining; 1, 1–10% positive cells; 2, 10–50% positive cells; 3, > 50% positive cells. the final score was evaluated by multiplying the staining intensity and stained area percentage. the negative and weak staining cases were considered as low expression group while the moderate and strong staining cases were defined as high expression group in later analysis.

Techniques: Gene Expression, Expressing

Journal: Future science OA

Article Title: Adverse predictive value of ASPM on lung adenocarcinoma overall survival depended on chemotherapy status.

doi: 10.1080/20565623.2025.2489328

Figure Lengend Snippet: Figure 2. (A) The representative immunohistochemical staining of ASPM negative, weak, moderate, and strong cases. (B) In our cohort, high ASPM expression predicted better LUAD prognosis (Log-rank test p = 5.4e-4). (C) In patients who received chemotherapy, high ASPM expression was related to better overall survival (Log-rank test p = 5.0e-4). (D) In patients who did not received chemotherapy, ASPM expression was not related to patient overall survival (Log-rank test p = 0.48). (E) Multivariate regression analysis showed ASPM was not an independent prognostic factor while other protective factor included chemother apy and risk factor included clinical stages. (F) time dependent ROC analysis was conducted at time points 365, 1095, and 1825 days. The AUC and confidence intervals were evaluated using the ci function of pROC.

Article Snippet: Immunohistochemistry and special staining the Luad tissues (n = 160) were obtained from surgically resected specimens and made into tissue microarray. the sections were de-paraffinized and hydrated, and the endogenous peroxidase was blocked. antigen retrieval was performed using the dako target Retrieval Solution, High pH (dako ominis, agilent technologies, Santa Clara, Ca, uSa), in a ptLink set at 98 °C for 25 min. the tissue sections were incubated with an anti-aSpm rabbit polyclonal antibody (1:200, a02584-1, Boster, China) 1h at 37 °C. detection of immunostaining was achieved by an enzyme-conjugated polymer complex (K8002, dako, denmark) adapted for autostainers from daKo (dako autostainer, agilent technologies). the staining intensity in selected areas was scored as follows: 0, no staining; 1, weak staining; 2, moderate staining; 3, strong staining. the stained area was scored as follows: 0, no staining; 1, 1–10% positive cells; 2, 10–50% positive cells; 3, > 50% positive cells. the final score was evaluated by multiplying the staining intensity and stained area percentage. the negative and weak staining cases were considered as low expression group while the moderate and strong staining cases were defined as high expression group in later analysis.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Future science OA

Article Title: Adverse predictive value of ASPM on lung adenocarcinoma overall survival depended on chemotherapy status.

doi: 10.1080/20565623.2025.2489328

Figure Lengend Snippet: Figure 3. (A) heatmap of signature proteins in ASPM low or high groups from proteomics dataset. (B) High ASPM1 related signature proteins were enriched mainly in cell cycle and mitosis processes by Metascape analysis. (C) ASPM positive group in proteomics dataset showed higher expression of proliferation related proteins mainly including AURKB, PLK1, KIFC3, EZH2,CDKN2A, BRD3, CASP2, MKI67, CDK1, and MDC1.

Article Snippet: Immunohistochemistry and special staining the Luad tissues (n = 160) were obtained from surgically resected specimens and made into tissue microarray. the sections were de-paraffinized and hydrated, and the endogenous peroxidase was blocked. antigen retrieval was performed using the dako target Retrieval Solution, High pH (dako ominis, agilent technologies, Santa Clara, Ca, uSa), in a ptLink set at 98 °C for 25 min. the tissue sections were incubated with an anti-aSpm rabbit polyclonal antibody (1:200, a02584-1, Boster, China) 1h at 37 °C. detection of immunostaining was achieved by an enzyme-conjugated polymer complex (K8002, dako, denmark) adapted for autostainers from daKo (dako autostainer, agilent technologies). the staining intensity in selected areas was scored as follows: 0, no staining; 1, weak staining; 2, moderate staining; 3, strong staining. the stained area was scored as follows: 0, no staining; 1, 1–10% positive cells; 2, 10–50% positive cells; 3, > 50% positive cells. the final score was evaluated by multiplying the staining intensity and stained area percentage. the negative and weak staining cases were considered as low expression group while the moderate and strong staining cases were defined as high expression group in later analysis.

Techniques: Expressing

Journal: Future science OA

Article Title: Adverse predictive value of ASPM on lung adenocarcinoma overall survival depended on chemotherapy status.

doi: 10.1080/20565623.2025.2489328

Figure Lengend Snippet: Figure 4. (A) Proteomics data analysis showed ASPM positive group had higher MKI67 expression. (B) Western blot analysis of A549 cells and H1299 cells confirmed effectiveness of ASPM transfection and in vitro overexpression. (C-D) Statistical anal ysis of western blot images. (E-F) CCK8 assay proved overexpression of ASPM promoted tumor cell proliferation in A549 and H1299 cells. (G-H) When treated with cisplatin, overexpression of ASPM enhanced the lethality on A549 and H1299 tumor cells. (P value *<0.05, **<0.01, ***<0.001).

Article Snippet: Immunohistochemistry and special staining the Luad tissues (n = 160) were obtained from surgically resected specimens and made into tissue microarray. the sections were de-paraffinized and hydrated, and the endogenous peroxidase was blocked. antigen retrieval was performed using the dako target Retrieval Solution, High pH (dako ominis, agilent technologies, Santa Clara, Ca, uSa), in a ptLink set at 98 °C for 25 min. the tissue sections were incubated with an anti-aSpm rabbit polyclonal antibody (1:200, a02584-1, Boster, China) 1h at 37 °C. detection of immunostaining was achieved by an enzyme-conjugated polymer complex (K8002, dako, denmark) adapted for autostainers from daKo (dako autostainer, agilent technologies). the staining intensity in selected areas was scored as follows: 0, no staining; 1, weak staining; 2, moderate staining; 3, strong staining. the stained area was scored as follows: 0, no staining; 1, 1–10% positive cells; 2, 10–50% positive cells; 3, > 50% positive cells. the final score was evaluated by multiplying the staining intensity and stained area percentage. the negative and weak staining cases were considered as low expression group while the moderate and strong staining cases were defined as high expression group in later analysis.

Techniques: Expressing, Western Blot, Transfection, In Vitro, Over Expression, CCK-8 Assay

Journal: Technology in Cancer Research & Treatment

Article Title: Cancer-Testis Gene Expression in Hepatocellular Carcinoma: Identification of Prognostic Markers and Potential Targets for Immunotherapy

doi: 10.1177/1533033820944274

Figure Lengend Snippet: Top 10 Hub Genes With Higher Degree Scores.

Article Snippet: Primary antibodies used for immunohistochemistry (IHC) of paraffin-embedded 72 HCC tissues and their paired noncancerous tissues included rabbit polyclonal anti-BUB1B antibody (1:200, Proteintech, Cat No. 11504-2-AP), rabbit polyclonal anti-ASPM antibody (1:100, Proteintech, Cat No. 26223-1-AP), rabbit polyclonal anti-beta catenin antibody (1:2000, Proteintech, Cat No. 51067-2-AP), and mouse monoclonal anti-CD8 antibody (1:200, Abcam, Cat No. ab17147).

Techniques: Protein Binding, Binding Assay, Activity Assay, Transduction, Ubiquitin Proteomics, Phospho-proteomics

Journal: Technology in Cancer Research & Treatment

Article Title: Cancer-Testis Gene Expression in Hepatocellular Carcinoma: Identification of Prognostic Markers and Potential Targets for Immunotherapy

doi: 10.1177/1533033820944274

Figure Lengend Snippet: Correlation between methylation levels and cancer-testis (CT) gene expression. Promoter methylation and gene expression data from The Cancer Genome Atlas (TCGA) were visualized using cBioPortal to demonstrate the correlation among selected hub CT genes, and significant negative relationship was identified in: (A) ARUKA, (B) ASPM, (C) BUB1, (D) BUB1B, (E) CCNB2, (F) NUF2, (G) PBK, (H) TPX2.

Article Snippet: Primary antibodies used for immunohistochemistry (IHC) of paraffin-embedded 72 HCC tissues and their paired noncancerous tissues included rabbit polyclonal anti-BUB1B antibody (1:200, Proteintech, Cat No. 11504-2-AP), rabbit polyclonal anti-ASPM antibody (1:100, Proteintech, Cat No. 26223-1-AP), rabbit polyclonal anti-beta catenin antibody (1:2000, Proteintech, Cat No. 51067-2-AP), and mouse monoclonal anti-CD8 antibody (1:200, Abcam, Cat No. ab17147).

Techniques: Methylation, Gene Expression

Journal: Technology in Cancer Research & Treatment

Article Title: Cancer-Testis Gene Expression in Hepatocellular Carcinoma: Identification of Prognostic Markers and Potential Targets for Immunotherapy

doi: 10.1177/1533033820944274

Figure Lengend Snippet: Prognostic values of 9 cancer-testis (CT) genes identified by overall survival analysis using Kaplan-Meier curve from GEPIA: (A) ASPM, (B) ARUKA, (C) BUB1, (D) BUB1B, (E) CDC45, (F) DLGAP5, (G) NUF2, (H) PBK, (I) TPX2.

Article Snippet: Primary antibodies used for immunohistochemistry (IHC) of paraffin-embedded 72 HCC tissues and their paired noncancerous tissues included rabbit polyclonal anti-BUB1B antibody (1:200, Proteintech, Cat No. 11504-2-AP), rabbit polyclonal anti-ASPM antibody (1:100, Proteintech, Cat No. 26223-1-AP), rabbit polyclonal anti-beta catenin antibody (1:2000, Proteintech, Cat No. 51067-2-AP), and mouse monoclonal anti-CD8 antibody (1:200, Abcam, Cat No. ab17147).

Techniques:

Journal: Human molecular genetics

Article Title: Kinetochore KMN network gene CASC5 mutated in primary microcephaly.

doi: 10.1093/hmg/dds386

Figure Lengend Snippet: Figure 2. Prioritization of candidate genes. (A) Global ranking of our candidate genes using the Endeavor program, higher ranking indicating more likely candidate. (B) Computation of the Ka/Ks ratios in primates (human versus macaque, first column) and in rodents (mouse versus rat, second column). Higher ratios reflect more rapid evolution. The third column is the ratio between Ka/Ks in primates and rodents. A ratio of .1 indicates accelerated evolution of the gene in the Homo Sapiens lineage. MCPH1, CDK5RAP2, CENPJ and ASPM are known MCPH genes. (C) Transcriptome study. Heatmap generated using the GenePattern software, com- paring expressions of all genes in the 2.7 Mb linkage interval, as well as CEP152, in control lymphoblasts and in lymphoblasts from patients E3 and S1. Red indicates the highest gene expression; dark blue indicates the lowest expression. Some transcripts are targeted by several probes indicated as a, b, c, d.

Article Snippet: Anti-CASC5 rabbit polyclonal antibody (Bethyl, catA300-805A; Abcam, ab70537), Anti-Blinkin mouse monoclonal antibody 31F2 (19), anti-ZWINT-1 rabbit polyclonal antibody (Abcam, ab84367), anti-BUBR1 mouse monoclonal antibody (Abcam, ab54894), anti-ASPM rabbit polyclonal antibody (Bethyl, catIHC-00058), anti-CEP152 rabbit polyclonal antibody (Bethyl, catA302-480A), anti-PCTN rabbit polyclonal antibody (Covance, pRb-432C) and anti-aTUBULIN mouse polyclonal (Sigma,T6793) were obtained commercially.

Techniques: Generated, Software, Control, Gene Expression, Expressing

Journal: Human molecular genetics

Article Title: Kinetochore KMN network gene CASC5 mutated in primary microcephaly.

doi: 10.1093/hmg/dds386

Figure Lengend Snippet: Figure 4. Functional effect of the mutation. (A) RT-PCR of random-primed RNA extracted from our patient’s lymphoblastoı¨d cell lines. Amplified cDNAs using primers surrounding CASC5 exon 18 were loaded on 6% acrylamide gel. F: Unaffected father (family E); P: MCPH patient E3; C1-C4: unrelated normal con- trols; RT+: positive and negative control of retrotranscription. Arrows indicate primers. A smaller band is observed in the homozygous patient and faintly in heterozygous father. Direct sequencing of this band extracted from the gel showed read-through from exon 17 into exon 19. Sequencing of the larger band at the expected size showed inclusion of exon 18 containing the mutation. (B) Ex vivo splicing assay for CASC5 variant. The cells were transfected with vectors con- taining two exons surrounding our exon of interest. One of the vectors contained the mutated exon 18 (EM), the other contained the wild-type exon 18 (EWT). Expressed mRNAs were detected on agarose gels. (C) Indicated amounts of whole cell lysates from HEK293T, HELA and HTB10 cells were separated on SDS-PAGE. CASC5 was detected by western blot using a commercial anti-CASC5 antibody. (D) Electrophoresis 100 mg of whole cell lysates from our patient’s lymphoblastoı¨d cell lines, and western blotting detection of CASC5 using a polyclonal antibody against customized CASC5 peptides (C-term: left; N-term: right). F: Unaffected father (family E); P: MCPH patient E3.

Article Snippet: Anti-CASC5 rabbit polyclonal antibody (Bethyl, catA300-805A; Abcam, ab70537), Anti-Blinkin mouse monoclonal antibody 31F2 (19), anti-ZWINT-1 rabbit polyclonal antibody (Abcam, ab84367), anti-BUBR1 mouse monoclonal antibody (Abcam, ab54894), anti-ASPM rabbit polyclonal antibody (Bethyl, catIHC-00058), anti-CEP152 rabbit polyclonal antibody (Bethyl, catA302-480A), anti-PCTN rabbit polyclonal antibody (Covance, pRb-432C) and anti-aTUBULIN mouse polyclonal (Sigma,T6793) were obtained commercially.

Techniques: Functional Assay, Mutagenesis, Reverse Transcription Polymerase Chain Reaction, Random Primed, Acrylamide Gel Assay, Negative Control, Sequencing, Ex Vivo, Splicing Assay, Variant Assay, Transfection, SDS Page, Western Blot, Electrophoresis