![Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 protein levels are elevated in human breast cancers. TMAs containing cores from invasive ductal carcinoma (IvDC), infiltrated ductal carcinoma (IfDC), intraductal carcinoma (IDC) and invasive lobular carcinoma (ILC) as well as pair-matched normal (PM-N) or cancer-free patients (CF-N) were analyzed for Spy1 expression. The Spy1 signal intensity was normalized to nuclear stain (TOTO-3/PI) signal. Patient numbers are indicated below the sample (N). Breast normal and cancer cell lines were analyzed by western blot analysis. Actin was used as a loading control. One representative blot of 2. Image collected and cropped by Citeab from the following publication (The cyclin-like protein Spy1/RINGO promotes mammary transformation and is elevated in human breast cancer. BMC Cancer (2012)) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Speedy-Ringo-Antibody-Western-Blot-NB100-2521-img0006.jpg "Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 protein levels are elevated in human breast cancers. TMAs containing cores from invasive ductal carcinoma (IvDC), infiltrated ductal carcinoma (IfDC), intraductal carcinoma (IDC) and invasive lobular carcinoma (ILC) as well as pair-matched normal (PM-N) or cancer-free patients (CF-N) were analyzed for Spy1 expression. The Spy1 signal intensity was normalized to nuclear stain (TOTO-3/PI) signal. Patient numbers are indicated below the sample (N). Breast normal and cancer cell lines were analyzed by western blot analysis. Actin was used as a loading control. One representative blot of 2. Image collected and cropped by Citeab from the following publication (The cyclin-like protein Spy1/RINGO promotes mammary transformation and is elevated in human breast cancer. BMC Cancer (2012)) licensed under a CC-BY license.")
| Reactivity | Hu, Mu, Rt, ShSpecies Glossary |
| Applications | WB, ICC/IF, IHC |
| Clonality | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| Format | BSA Free |
| Concentration | 1.1 mg/ml |
| Immunogen | A synthetic peptide made to a C-terminal region of human Speedy/Ringo (within residues 200-286). This immunogen is conserved in both isoforms 1 and 2. [Swiss-Prot Q5MJ69] |
| Localization | Nuclear |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Rabbit |
| Gene | SPDYA |
| Purity | Immunogen affinity purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Dilutions |
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| Application Notes | In Western blot analysis, a band is seen at ~36 kDa (isoform 2). After a longer exposure, a 31 kDa band can be detected (isoform 1). There is also a non-specific band at ~58 kDa. |
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| Storage | Store at 4C. Do not freeze. |
| Buffer | Tris-Citrate/Phosphate (pH 7.0 - 8.0) |
| Preservative | 0.1% Sodium Azide |
| Concentration | 1.1 mg/ml |
| Purity | Immunogen affinity purified |
Secondary Antibodies |
Isotype Controls |
Research Areas for Speedy/Ringo Antibody (NB100-2521)Find related products by research area.
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The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.
| Gene Symbol | SPDYA |
![Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 stable protein induces anchorage independent growth. Representative foci after 14 day soft agar assay visualized using light microscopy. Ras-V12 is transfected as a positive control. n = 3. Total numbers of foci were counted over 3 separate plates using separate transfections for each experiment. n = 3. Error bars reflect SE between triplicate experiments. t test was performed. Western blot analysis of experiments performed. Quantified using densitometry followed by normalization for Actin levels. Image collected and cropped by Citeab from the following publication (The cyclin-like protein Spy1/RINGO promotes mammary transformation and is elevated in human breast cancer. BMC Cancer (2012)) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Speedy-Ringo-Antibody-Western-Blot-NB100-2521-img0005.jpg "Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 stable protein induces anchorage independent growth. Representative foci after 14 day soft agar assay visualized using light microscopy. Ras-V12 is transfected as a positive control. n = 3. Total numbers of foci were counted over 3 separate plates using separate transfections for each experiment. n = 3. Error bars reflect SE between triplicate experiments. t test was performed. Western blot analysis of experiments performed. Quantified using densitometry followed by normalization for Actin levels. Image collected and cropped by Citeab from the following publication (The cyclin-like protein Spy1/RINGO promotes mammary transformation and is elevated in human breast cancer. BMC Cancer (2012)) licensed under a CC-BY license.")
![Immunohistochemistry-Paraffin: Speedy/Ringo Antibody [NB100-2521] - Speedy/Ringo was detected in immersion fixed paraffin-embedded sections of human testis using Rabbit Anti-Human Speedy/Ringo polyclonal Antibody (Catalog # NB100-2521) at 1:300 for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nuclei of sperm cells.](http://images.novusbio.com/fullsize/Speedy-Ringo-Antibody-Immunohistochemistry-Paraffin-NB100-2521-img0004.jpg "Immunohistochemistry-Paraffin: Speedy/Ringo Antibody [NB100-2521] - Speedy/Ringo was detected in immersion fixed paraffin-embedded sections of human testis using Rabbit Anti-Human Speedy/Ringo polyclonal Antibody (Catalog # NB100-2521) at 1:300 for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nuclei of sperm cells.")
![Western Blot: Speedy/Ringo Antibody [NB100-2521] - Detection of Speedy/Ringo in mouse testis (2ug/ml).](http://images.novusbio.com/fullsize/Speedy-Ringo-Antibody-Western-Blot-NB100-2521-img0003.jpg "Western Blot: Speedy/Ringo Antibody [NB100-2521] - Detection of Speedy/Ringo in mouse testis (2ug/ml).")
![Western Blot: Speedy/Ringo Antibody [NB100-2521] - Elevated Spy1 levels prevent contact inhibition. (A) Representative views of focus formation assays in NIH3T3 cells transfected with Myc-Spy1-WT, Myc-Spy1-TST, positive control Ras-V12 or empty vector PCS3 control. n = 3 (B) Quantification of the number of colonies. Error bars reflect SE between triplicate experiments. t test was performed;** P ≤ 0.01. n = 3 (right panel) Western blot analysis of one representative lysate from A-B. Image collected & cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-2521_rabbit-polyclonal-speedy-ringo-antibody-31020241534341.jpg "Western Blot: Speedy/Ringo Antibody [NB100-2521] - Elevated Spy1 levels prevent contact inhibition. (A) Representative views of focus formation assays in NIH3T3 cells transfected with Myc-Spy1-WT, Myc-Spy1-TST, positive control Ras-V12 or empty vector PCS3 control. n = 3 (B) Quantification of the number of colonies. Error bars reflect SE between triplicate experiments. t test was performed;** P ≤ 0.01. n = 3 (right panel) Western blot analysis of one representative lysate from A-B. Image collected & cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 stable protein induces anchorage independent growth. (A) 293 cells were transfected with different concentrations of Myc-Spy1-WT, Myc-Spy1-TST or 30 μg of empty PCS3 vector control. Soft agar assay was carried out & plates photographed & quantified on day 14. Foci were averaged over 3 separate transfections for each experiment. n = 3 (B) Western blot analysis of one representative experiment; densitometry in lower panel. n = 3 (C) Representative foci after 14 day soft agar assay visualized using light microscopy. Ras-V12 is transfected as a positive control. n = 3. (D) Total numbers of foci were counted over 3 separate plates using separate transfections for each experiment. n = 3. Error bars reflect SE between triplicate experiments. t test was performed;** P ≤ 0.01. (E) Western blot analysis of experiments in Figure. 1C & D. Quantified using densitometry followed by normalization for Actin levels. (A-E) Error bars reflect SE between triplicate experiments. t test was performed;* P≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001. Image collected & cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-2521_rabbit-polyclonal-speedy-ringo-antibody-310202415541987.jpg "Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 stable protein induces anchorage independent growth. (A) 293 cells were transfected with different concentrations of Myc-Spy1-WT, Myc-Spy1-TST or 30 μg of empty PCS3 vector control. Soft agar assay was carried out & plates photographed & quantified on day 14. Foci were averaged over 3 separate transfections for each experiment. n = 3 (B) Western blot analysis of one representative experiment; densitometry in lower panel. n = 3 (C) Representative foci after 14 day soft agar assay visualized using light microscopy. Ras-V12 is transfected as a positive control. n = 3. (D) Total numbers of foci were counted over 3 separate plates using separate transfections for each experiment. n = 3. Error bars reflect SE between triplicate experiments. t test was performed;** P ≤ 0.01. (E) Western blot analysis of experiments in Figure. 1C & D. Quantified using densitometry followed by normalization for Actin levels. (A-E) Error bars reflect SE between triplicate experiments. t test was performed;* P≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001. Image collected & cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Immunocytochemistry/ Immunofluorescence: Speedy/Ringo Antibody [NB100-2521] - CDC2 associates with SPDY at ERES. (A-C) 0 h GV stage oocyte labeled for CDC2 (A, green), DNA (A, blue), & SPDY (B, red). Both CDC2 & SPDY localize to the same cortical domain (C). Note that the center of this oocyte is dented (the area that contains the arrow) causing the structure to appear in the middle of the oocyte, whereas it is located in the cortex. (D-F) 0 h GV stage oocyte stained for Cyclin B (D, green), DNA (D, blue), & P-GM130 (E, red). Cyclin B did not localize to the P-GM130-labeled cortical domain (F). (G-I) 0 h GV stage oocyte labeled for PSTAIR (G, green), DNA (G, blue), & P-GM130 (H, red). Spatial overlap of PSTAIR & phosphorylated GM130 staining (yellow) in a cortical domain is evident in the merged image (I). Images are Z-projections of 2 (A-C) or 3 (D-I) consecutive sections; scale bars represent 20 μm in C, F, & I, & 5 μm in C', F', & I'. Arrows indicate the region of the oocyte that is shown enlarged in the insets (A'-I'). Arrowheads denote the position of the GV. Image collected & cropped by CiteAb from the following publication (//bmcdevbiol.biomedcentral.com/articles/10.1186/1471-213X-9-8), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-2521_rabbit-polyclonal-speedy-ringo-antibody-31020241555219.jpg "Immunocytochemistry/ Immunofluorescence: Speedy/Ringo Antibody [NB100-2521] - CDC2 associates with SPDY at ERES. (A-C) 0 h GV stage oocyte labeled for CDC2 (A, green), DNA (A, blue), & SPDY (B, red). Both CDC2 & SPDY localize to the same cortical domain (C). Note that the center of this oocyte is dented (the area that contains the arrow) causing the structure to appear in the middle of the oocyte, whereas it is located in the cortex. (D-F) 0 h GV stage oocyte stained for Cyclin B (D, green), DNA (D, blue), & P-GM130 (E, red). Cyclin B did not localize to the P-GM130-labeled cortical domain (F). (G-I) 0 h GV stage oocyte labeled for PSTAIR (G, green), DNA (G, blue), & P-GM130 (H, red). Spatial overlap of PSTAIR & phosphorylated GM130 staining (yellow) in a cortical domain is evident in the merged image (I). Images are Z-projections of 2 (A-C) or 3 (D-I) consecutive sections; scale bars represent 20 μm in C, F, & I, & 5 μm in C', F', & I'. Arrows indicate the region of the oocyte that is shown enlarged in the insets (A'-I'). Arrowheads denote the position of the GV. Image collected & cropped by CiteAb from the following publication (//bmcdevbiol.biomedcentral.com/articles/10.1186/1471-213X-9-8), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 stable protein accelerates tumorigenesis in vivo.(A) Percentage of mice presenting with palpable tumors from 0-19 days post-transplant. Each data point represents 4 mice per indicated construct. The entire experiment was repeated three times using three independently derived overexpressing cell lines for each construct. Mann-Whitney Test was performed (p < 0.05). (A; lower blot) Western blots were conducted to measure the stability of Myc-Spy1-WT (left) & Myc-Spy1-TST (right) from representative time points. Empty vector control (Cntl) cell expression levels of Spy1 are seen in lane 1 of each blot. (B) Total tumor volume was calculated for both Spy1-HC11 (Spy1-WT) & Spy1-TST-HC11 (Spy1-TST) transplanted glands. Results were taken from 45 transplants using cells from 3 separate transfections. Error bars reflect SE between transplants from different transfections. Left hand panel reflects overall volume, right hand panel reflects volumes of tissues taken over a month post-transplant. Image collected & cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-2521_rabbit-polyclonal-speedy-ringo-antibody-31020241553735.jpg "Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 stable protein accelerates tumorigenesis in vivo.(A) Percentage of mice presenting with palpable tumors from 0-19 days post-transplant. Each data point represents 4 mice per indicated construct. The entire experiment was repeated three times using three independently derived overexpressing cell lines for each construct. Mann-Whitney Test was performed (p < 0.05). (A; lower blot) Western blots were conducted to measure the stability of Myc-Spy1-WT (left) & Myc-Spy1-TST (right) from representative time points. Empty vector control (Cntl) cell expression levels of Spy1 are seen in lane 1 of each blot. (B) Total tumor volume was calculated for both Spy1-HC11 (Spy1-WT) & Spy1-TST-HC11 (Spy1-TST) transplanted glands. Results were taken from 45 transplants using cells from 3 separate transfections. Error bars reflect SE between transplants from different transfections. Left hand panel reflects overall volume, right hand panel reflects volumes of tissues taken over a month post-transplant. Image collected & cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 protein levels are elevated in human breast cancers.(A) TMAs containing cores from invasive ductal carcinoma (IvDC), infiltrated ductal carcinoma (IfDC), intraductal carcinoma (IDC) & invasive lobular carcinoma (ILC) as well as pair-matched normal (PM-N) or cancer-free patients (CF-N) were analyzed for Spy1 expression. The Spy1 signal intensity was normalized to nuclear stain (TOTO-3/PI) signal. Patient numbers are indicated below the sample (N). (B) Breast normal & cancer cell lines were analyzed by western blot analysis. Actin was used as a loading control. One representative blot of 2. (C) Western blot of 3 replicate infections of pLKO control or pLKO Spy1 in MDA-231 cells. Middle panel represents the densitometry values over 3 separate experiments. Where Spy1 is corrected for actin levels. Lower panel reflects cell counts at 4, 6, & 24 h post-infection. (All panels) Data shown is mean ± s.d. Student's t-test was performed * P < 0.05;**P < 0.01. (D) Knockdown effects on cell counts of MCF7 cells over 72 h after transfection with either pSUPER empty vector (siCntl) or pSUPER-Spy1 (siSpy1) over 3 separate transfections. Data shown is mean ± s.d. Image collected & cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-2521_rabbit-polyclonal-speedy-ringo-antibody-31020241555227.jpg "Western Blot: Speedy/Ringo Antibody [NB100-2521] - Spy1 protein levels are elevated in human breast cancers.(A) TMAs containing cores from invasive ductal carcinoma (IvDC), infiltrated ductal carcinoma (IfDC), intraductal carcinoma (IDC) & invasive lobular carcinoma (ILC) as well as pair-matched normal (PM-N) or cancer-free patients (CF-N) were analyzed for Spy1 expression. The Spy1 signal intensity was normalized to nuclear stain (TOTO-3/PI) signal. Patient numbers are indicated below the sample (N). (B) Breast normal & cancer cell lines were analyzed by western blot analysis. Actin was used as a loading control. One representative blot of 2. (C) Western blot of 3 replicate infections of pLKO control or pLKO Spy1 in MDA-231 cells. Middle panel represents the densitometry values over 3 separate experiments. Where Spy1 is corrected for actin levels. Lower panel reflects cell counts at 4, 6, & 24 h post-infection. (All panels) Data shown is mean ± s.d. Student's t-test was performed * P < 0.05;**P < 0.01. (D) Knockdown effects on cell counts of MCF7 cells over 72 h after transfection with either pSUPER empty vector (siCntl) or pSUPER-Spy1 (siSpy1) over 3 separate transfections. Data shown is mean ± s.d. Image collected & cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
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![Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).](https://images.novusbio.com/images/Goat-anti-Rabbit-IgG-H+L-Secondary-Antibody-HRP-Western-Blot-NB7160-img0001.jpg "Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).")
![Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.](https://images.novusbio.com/images/Rabbit--Mouse-IgG-Isotype-Control-Flow-Cytometry-NBP2-24891-img0001.jpg "Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.")