rabbit anti ncx2  (Bioss)

Journal: International Journal of Molecular Sciences

Article Title: Hamster Sperm Possess Functional Na + /Ca 2+ -Exchanger 1: Its Implication in Hyperactivation

doi: 10.3390/ijms24108905

Figure Lengend Snippet: NCX isoforms detected by RNA-seq analyses from hamster testes. NCX genes detected by RNA-seq analysis and bit score, e-value, % of identity, TPM (transcripts per kilobase million) and FPKM (fragments per kilobase of exon per million mapped reads) values were summarized.

Article Snippet: The custom rabbit anti-NCX2 antiserum was obtained by immunizing rabbits with the peptide (CIGAEGDPPKSIELD) conjugated with Keyhole Limpet Hemocyanin (Merck KgaA, Darmstadt, Germany).

Techniques: Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Hamster Sperm Possess Functional Na + /Ca 2+ -Exchanger 1: Its Implication in Hyperactivation

doi: 10.3390/ijms24108905

Figure Lengend Snippet: The expression of NCXs in hamster spermatozoa. The expression of NCX1 and NCX2 was examined via RT–PCR using hamster testis total RNA ( A ), and Western blotting using microsomal protein preparations from hamster spermatozoa ( B – D ). ( A ) The expression of NCX1 and NCX2 mRNA in hamster testes was investigated using conventional RT–PCR. PCR products without templates were used as the negative control. ( B ) The expression of NCX1 protein in hamster spermatozoa was investigated via Western blotting using two different antibodies (D3F3H from CST and R3F1 from Swant) against NCX1. ( C ) The R3F1 antibody was preabsorbed by blocking peptide, and Western blotting was performed to validate the specificity of the antibody. ( D ) The expression of NCX2 protein in hamster spermatozoa and testes were investigated using custom rabbit anti–NCX2 antiserum. Hamster brain samples were used as a positive control. The results of antiserum (left) and preimmune serum (right) are shown side by side. The amount of protein loaded was 10 µg/well. Red arrowheads indicate a positive band. The molecular weights in kDa are shown on the left side.

Article Snippet: The custom rabbit anti-NCX2 antiserum was obtained by immunizing rabbits with the peptide (CIGAEGDPPKSIELD) conjugated with Keyhole Limpet Hemocyanin (Merck KgaA, Darmstadt, Germany).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Negative Control, Blocking Assay, Positive Control

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: Downregulation of NCX protein expression in colon cancer (CC). a Western blotting was used to compare the expression of the three isoforms of NCXs in tumor tissues with that in the normal colon tissue of 111 CC patients. The intensity of each band was analyzed. The NCX1 ( b ), NCX2 ( c ), and NCX3 ( c ) protein levels in tumors, in general, were significantly lower than those in normal tissue, particularly those of NCX1 and NCX3. The ratio of NCX to GAPDH is shown by a histogram. (T: colonic malignant tumor; N: normal colonic tissue; paired t test; * p < 0.05 and ** p < 0.01 compared with T)

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques: Expressing, Western Blot

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: NCX mRNA expression was decreased in CC. a – c In 111 patients, the overall expression of NCX mRNA in most tumor samples was lower than that in normal tissues by qPCR. Compared with the three isoforms, the mRNA of NCX1 ( a ) and NCX3 ( c ) in tumors decreased more remarkably than that of NCX2 ( b ). Histograms representing the relative expression of NCX compared to GAPDH (paired t test; * p < 0.05, ** p < 0.01)

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques: Expressing

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: Expression of NCX1, NCX2, and NCX3 in normal and tumor tissues of CC by HE and IHC stainings (×200). a HE staining for normal colonic mucosa. The structure of normal colonic mucosa. The glands were arranged in order. The nuclei of epithelial cells and stromal cells were clear. b In the normal colon, NCX1 was highly expressed in the cell membrane of normal colonic glandular epithelial cells and stromal cells in the mucosa, and the closer to the intestinal cavity, the stronger the staining was. c NCX2 was moderately expressed in stromal cells and glandular epithelium. d The expression of NCX3 mainly in stromal cells and submucosa. e HE staining for colonic malignant glands. The glands showed disorganized malignant growth. The nucleus was large and hyperchromatic with vacuoles. f NCX1 was moderately expressed in malignant glands. g The positive expression of NCX2 was noticeably decreased compared to that in normal tissues. h The staining intensity of the NCX3 protein in tumor tissue was very low by IHC. i The integral optical density (IOD) is represented by a histogram. (tumor: colonic malignant tumor; normal: normal colonic tissue; black arrows refer to glands, white arrows refer to stromal cells; one-way ANOVA; * p < 0.05 and ** p < 0.01, compared with NCX1-tumor; # p < 0.05, compared with NCX2-tumor; @ p < 0.05, compared with NCX3-tumor)

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques: Expressing, Staining

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: Expression of NCX1, NCX2, and NCX3 in different immune cells of the normal colon (×400). a Some macrophages (black dovetail arrows) were found in the submucosa of the normal colon. b The protein expression strength of NCX1 in macrophages was medium but weaker than that of NCX2 ( c ). d Intense expression of NCX3 in macrophages. e In lymphoid follicles of the normal colon, B cells and T cells are indicated by black arrows and white arrows, respectively. f The expression of NCX1 was intense in lymphocytes (B cells and T cells). g NCX2 was mainly moderately expressed in lymphocytes. h NCX3 was weakly expressed in lymphocytes

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques: Expressing

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: Expression of NCX1, NCX2, and NCX3 in primary and metastatic sites. (×400). a – c In the primary tumor site, heteromorphism of the glands was very obvious, and the degree of differentiation was moderate, as determined. The expression strengths of NCX1 ( a ), NCX2 ( b ), and NCX3 ( c ) in colonic malignant glands decreased more obviously. d – f The expression intensity of NCXs at lymph node metastasis sites decreased more significantly than that at primary sites. d The expression of NCX1 was weak in malignant glands compared with lymph node metastasis sites. There were nearly no positive NCX2 ( e ) and NCX3 ( f ) cells at the lymph node metastasis sites. g – i The expression of NCXs in normal hepatocytes (black arrows) at liver metastasis sites was obvious, and the strength of NCX1 staining ( g ; white arrow) was higher than that of NCX2 ( h ; white arrow) and NCX3 ( i ; white arrow). NCXs were almost not expressed in malignant glands ( g , h , i ; white arrows). The IOD is represented by a histogram ( j – l )

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques: Expressing, Staining

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: Correlation between NCXs and the clinicopathological features of colon cancer

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques: Expressing

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: Decreased NCX expression was negatively correlated with the overall survival (OS) time. a Kaplan-Meier curves of the OS of 111 CC patients are displayed according to the NCX1 low and NCX1 high expression groups. b Kaplan-Meier survival analysis curves are also displayed for different NCX2 expression groups. c Kaplan-Meier survival analysis curves are also displayed for different NCX3 expression groups. The log-rank test indicated a significant difference between the survival curves (** p < 0.01; ## p < 0.01; @@ p < 0.01)

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques: Expressing

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: Decreased expression of NCXs was negatively correlated with the disease-free survival (DFS) time. a Kaplan-Meier curves of the DFS of 92 preoperative non-metastatic CC patients are displayed according to the NCX1 low and NCX1 high expression groups. b Kaplan-Meier survival analysis curves are also displayed for different NCX2 expression groups. c Kaplan-Meier survival analysis curves are also displayed for different NCX3 expression groups. The log-rank test indicated a significant difference between the survival curves (** p < 0.01; ## p < 0.01; @@ p < 0.01)

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques: Expressing

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: Cox univariate and multivariate survival analyses to analyze the independent prognostic factors for colon cancer

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques:

Journal: World Journal of Surgical Oncology

Article Title: The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

doi: 10.1186/s12957-021-02284-5

Figure Lengend Snippet: NCXs can predict the prognosis of CC. According to the grayscale value of NCX expression in tumor tissue (tumor) and matched normal tissue (normal) calculated by WB bands, the ratios (tumor/normal) of NCX were obtained. Three group ratios of NCX1, NCX2, and NCX3 ( n = 111; NCXs were identified by different curves) were used to draw ROC curves. The ROC curves used the NCX (NCX1, NCX, and NCX3) ratios to define the NCX cutoff values related to OS. The ROC curves of NCX1, NCX2, and NCX3 were used to predict the OS of 111 CC patients. The abscissa represents sensitivity, and the ordinate represents specificity. Numerical analysis of the area under the ROC curve showed significant differences

Article Snippet: The sections were incubated with mouse anti-NCX1 monoclonal antibody (1:400, Abcam, Cambridge, UK), rabbit anti-NCX2 polyclonal antibody (1: 200, Bioss, Beijing, P.R.

Techniques: Expressing