Rabbit monoclonal [EPR1574] to Stathmin 1 (phospho S63)
Specificityab76583 only detects Stathmin 1 phosphorylated on Serine 62.
The antibody immunogen shares 86% homology with Stathmin-2, therefore it is possible that the antibody will cross-react with Stathmin-2 when phosphorylated at serine 97. This has not been assessed experimentally.
HeLa cell lysates treated with Calyculin A; human brain tissue.
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated "PUR" on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
Application notesIs unsuitable for Flow Cyt or ICC.
FunctionInvolved in the regulation of the microtubule (MT) filament system by destabilizing microtubules. Prevents assembly and promotes disassembly of microtubules. Phosphorylation at Ser-16 may be required for axon formation during neurogenesis. Involved in the control of the learned and innate fear.
Tissue specificityUbiquitous. Expression is strongest in fetal and adult brain, spinal cord, and cerebellum, followed by thymus, bone marrow, testis, and fetal liver. Expression is intermediate in colon, ovary, placenta, uterus, and trachea, and is readily detected at substantially lower levels in all other tissues examined. Lowest expression is found in adult liver. Present in much greater abundance in cells from patients with acute leukemia of different subtypes than in normal peripheral blood lymphocytes, non-leukemic proliferating lymphoid cells, bone marrow cells, or cells from patients with chronic lymphoid or myeloid leukemia.
Sequence similaritiesBelongs to the stathmin family. Contains 1 SLD (stathmin-like) domain.
Post-translational modificationsMany different phosphorylated forms are observed depending on specific combinations among the sites which can be phosphorylated. MAPK is responsible for the phosphorylation of stathmin in response to NGF. Phosphorylation at Ser-16 seems to be required for neuron polarization (By similarity). Phosphorylation at Ser-63 reduces tubulin binding 10-fold and suppresses the MT polymerization inhibition activity.
Western blot - Anti-Stathmin 1 (phospho S63) antibody [EPR1574] (ab76583)
All lanes : Anti-Stathmin 1 (phospho S63) antibody [EPR1574] (ab76583) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with nocodazole at 100 ng/mL for 18 hours. Whole cell lysates Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with nocodazole at 100 ng/mL for 18 hours. Whole cell lysates. Then the membrane was incubated with phosphatase.
Lysates/proteins at 15 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size : 17 kDa Observed band size : 17 kDa
Exposure time : 30 seconds
Blocking/Diluting buffer and concentration 2% BSA/TBST
ab76583, at a 1/250 dilution, staining Stathmin 1 in paraffin embedded human brain tissue by Immunohistochemistry.
Western blot - Stathmin 1 (phospho S62) antibody [EPR1574] (ab76583)
All lanes : Anti-Stathmin 1 (phospho S63) antibody [EPR1574] (ab76583) at 1/10000 dilution
Lane 1 : HeLa cell lysate, untreated Lane 2 : HeLa cell lysate treated with Calyculin A
Lysates/proteins at 10 µg per lane.
Secondary HRP labelled goat anti rabbit at 1/2000 dilution
Predicted band size : 17 kDa Observed band size : 17 kDa
Dot Blot - Anti-Stathmin 1 (phospho S63) antibody [EPR1574] (ab76583)
Dot blot analysis of Stathmin 1 (phospho S63) phospho peptide (Lane 1) and Stathmin 1 non-phospho peptide (Lane 2) labeling Stathmin 1 (phospho S63) with ab76583 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) at 1/100 000 was used as the secondary antibody.
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Western blot - Anti-Stathmin 1 (phospho S63) antibody [EPR1574] (ab76583)Image courtesy of Qifeng Lin by Abreview.
Predicted band size : 17 kDa
Image courtesy of Qifeng Lin by Abreview.
Alkaline phosphatase treatment removes S63 phosphorylation.
Samples treated with phosphatase were run alongside the normal lysate, and the phosph-S63 signal is not detected after phosphatase treatment, thus suggesting the signal is very specific to the phosphorylated S63. Whole cell lysate prepared from a human colon cancer cell line was loaded at 20µg.ab76583 used at a 1/1000 dilution. Secondary used was an HRP conjugated goat anti-rabbit polyclonal used at a 1/10000 dilution.
References for Anti-Stathmin 1 (phospho S63) antibody [EPR1574] (ab76583)
This product has been referenced in:
Yip YY et al. cAMP-dependent protein kinase and c-Jun N-terminal kinase mediate stathmin phosphorylation for the maintenance of interphase microtubules during osmotic stress. J Biol Chem289:2157-69 (2014).
Read more (PubMed: 24302736) »
Tian X et al. Microtubule dynamics control HGF-induced lung endothelial barrier enhancement. PLoS One9:e105912 (2014).
Read more (PubMed: 25198505) »