Catalog # SP3901

Slingshot-1L (Ser-978), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
ICC	1:100
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms, Ck

MW 150 kDa

Members of the ADF/cofilin (AC) family are actin-severing proteins that regulate actin remodeling during cell motility. Regulation of cofilin activity can occur through serine phosphorylation and dephosphorylation. Activation of cofilin kinases, LIMK1 or LIMK2, leads to phosphorylation of cofilin at serine 3. This phosphorylation disrupts cofilin binding to actin in vitro and in vivo. Multiple phosphatases, Slingshot, PP1, PP2A, PP2B, and chronophin can dephosphorylate Ser-3 and activate actin binding. In mammals, the Slingshot family includes SSH1L, SSH2L, and SSH3L. SSH1L and SSH2L mRNAs are widely expressed, while SSH3L has high expression in epithelial tissues. SSH1L can associate with F-actin and may be the major phosphatase regulating cofilin activity. Disruption of SSH1L expression using RNA interference impairs directional cell migration. Phosphorylation of SSH1L at Ser-937 and Ser-978 by PKD leads to association with 14-3-3, sequestration of the phosphatase to the cytoplasm, and reduces cell migration. Thus, Slingshot phosphatases may be critical for regulating cytoskeletal protein activity and cell motility.

References

Wang, Y. et al. (2015) Cancer Lett. 360(2):171 (ICC:PC cell lines, KLM1, KLM1-R, AsPC-1, BxPC-3, PK45p, PK59)
DiPaolo, B.C. et al. (2013) Am Jou of Phys 305(2): L141. (WB: rat alveolar type II)
Soosairajah, J. et al. (2005) EMBO J. 24(3):473.
Endo, M. et al. (2003) J Neurosci. 23(7):2527.
Niwa, R. et al. (2002) Cell 108:233.

Western blot of human recombinant SSH1L untreated (lanes 1 & 3) or treated with lambda phosphatase (lanes 2 & 4) and rat PC12 cells unstimulated (lanes 5 & 7) or stimulated with calyculin A (lanes 6 & 8). The blots were probed with anti-SSH1L (C-term.) (lanes 1, 2, 5, & 6) or anti-SSH1L (Ser-978) (lanes 3, 4, 7, & 8).

Immunocytochemical labeling of Slingshot-1L in rat PC12 cells differentiated with NGF. The cells were labeled with rabbit polyclonal anti-SSH1L (C-term.) and anti-SSH1L (Ser-978) antibodies, then detected using appropriate secondary antibody conjugated to Cy3 (Right panel). Phase image of corresponding PC12 cells (Left panel).

Slinshot-1L (Ser-978) antibody was generated from a phospho-peptide that included amino acids surrounding serine 978 in human SSH1L. This sequence has high homology to the conserved site in rat, mouse, chicken, and zebrafish SSH1L. The sequence also has high homology to the conserved site in SSH2L (Ser-1259).

*For more information, see UniProt Accession Q8WYL5

Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody was affinity purified using phospho-SSH1L (Ser-978) peptide (without carrier). The antibody detects a 150 kDa* full length recombinant human SSH1L phosphoprotein and this reactivity is eliminated with lambda phosphatase treatment. The antibody also detects SSH1L on immunoblots of rat PC12 and mouse C2C12 cells that were stimulated with calyculin A.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

Tedeschi, A. et al. (2019) Neuron. Online Aug. 7. (WB: rat DRG neurons)
Kermath, BA et al. (2019) Cereb Cortex. Apr 11. pii: bhz070. (WB: human brain)
Tan, F. et al. (2015) J Exp Clin Cancer Res. 9(34):117 (WB: HCT-116, LoVo, CRC tissues)
Bencsik, N. et al. (2015) J Cell Biol. 210(5):771. (WB: hippocampal dendritic spines)
Xiu, L. et al. (2015) Am J Pathol. 185(2):387 (WB: human MSCs and HPCs)
Barone, E. et al. (2014) Bioch Biophys Acta 1842(12A):2500. (WB: mouse brain)
Xiang, S.Y. et al. (2013) Sci Signal. 6(306):ra108. (WB: rat cardiomyocytes, S1P treated)
DiPaolo, B.C. et al. (2013) Am Jou of Phys 305(2): L141 (WB: rat alveolar type II)
Wang, Y. et al. (2013) J Neurosci. 33(15):6423. (WB: rat infralimbic cortex)
Vitolo, M. et al. (2013) Oncogene. 32(17):2200. (WB: MCF-10A, 293T)
Sparrow, N. et al. (2012) J Neurosci. 32(15):5284. (WB & ICC: rat Schwann cells)

This kit contains: