Speedy/Ringo Antibody

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.

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.

Western Blot: Speedy/Ringo Antibody [NB100-2521] - Detection of Speedy/Ringo in mouse testis (2ug/ml).

• Reactivity: Hu, Mu, Rt, Sh
• Applications: WB, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.1 mg/ml

Speedy/Ringo Antibody Summary

• 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

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/Immunofluorescence reported in scientific literature (PMID 19187565)
  • Immunohistochemistry reported in scientific literature (PMID 19187565)
  • Immunohistochemistry-Paraffin
  • Western Blot 2 ug/ml
• 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.
• Control:

  Mouse Testis Whole Tissue Lysate (Adult Whole Normal)

• Control Peptide:

  Speedy/Ringo Peptide (NB100-2521PEP)

Reactivity Notes

Sheep reactivity reported in scientific literature (PMID: 19187565). Human reactivity reported in scientific literature (PMID: 24037419). Rat reactivity reported in scientific literature (PMID: 19075091).

Packaging, Storage & Formulations

• 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

Alternate Names for Speedy/Ringo Antibody

• hSpy/Ringo A
• MGC110856
• Rapid inducer of G2/M progression in oocytes A
• RINGO A
• RINGO3
• SPDY1
• speedy homolog 1 (Drosophila)
• speedy homolog A (Xenopus laevis)
• speedy protein A
• speedy-1
• Spy1
• SPY1MGC57218

Background

Speedy/Ringo (Spy1) regulates the G1/S phase transition of the cell cycle by binding and activating CDC2, CDK2 and CDKN1B/KIP1. It mediates cell survival during the DNA damage process through activation of CDK2.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Speedy/Ringo Antibody (NB100-2521)(7)

Publications using NB100-2521

• Wang X, Zhu M, Shan D et al. Spy1, a Unique Cell Cycle Regulator, Alters Viability in ALS Motor Neurons and Cell Lines in Response to Mutant SOD1-Induced DNA Damage DNA Repair 2018-12-01 [PMID: 30630676] (WB)
• Fei M, Hang Q, Hou S, Ruan C. Cell adhesion to fibronectin down-regulates the expression of Spy1 and contributes to drug resistance in multiple myeloma cells. Int J Hematol. 2013-10-01 [PMID: 24037419] (WB, Human)
• Al Sorkhy M, Ferraiuolo R-M, Jalili E et al. The cyclin-like protein Spy1/RINGO promotes mammary transformation and is elevated in human breast cancer. BMC Cancer 2612(1):45. 2012-01-01 [PMID: 22280365]
• Holzenspies JJ, Stoorvogel W, Colenbrander B et al. CDC2/SPDY transiently associates with endoplasmic reticulum exit sites during oocyte maturation. BMC Dev Biol 2009-02-03 [PMID: 19187565] (IF/IHC, ICC/IF, Sheep)
• Ng LF, Chow A, Sun YJ et al. IL-1beta, IL-6, and RANTES as biomarkers of Chikungunya severity. PLoS One 2009-01-01 [PMID: 19156204]
• Giachini FR, Chiao CW, Carneiro FS et al. Increased activation of stromal interaction molecule-1/Orai-1 in aorta from hypertensive rats: a novel insight into vascular dysfunction. Hypertension 2009-02-01 [PMID: 19075091] (Rat)
• Al Sorkhy M et al. The cyclin-dependent kinase activator, Spy1A, is targeted for degradation by the ubiquitin ligase NEDD4. J Biol Chem;284(5):2617-27. 2009-01-30 [PMID: 19054764]

Bioinformatics

• Gene Symbol: SPDYA