Overview

  • Product name
    Anti-Stathmin 1 (phospho S16) antibody
    See all Stathmin 1 primary antibodies
  • Description
    Rabbit polyclonal to Stathmin 1 (phospho S16)
  • Host species
    Rabbit
  • Specificity
    This antibody detects endogenous levels of Stathmin 1 only when phosphorylated at serine 16. (Human: Ser16; Mouse: Ser16; Rat: Ser16).
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-P, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Stathmin 1. synthesized phosphopeptide derived from human Stathmin 1 around the phosphorylation site of serine 16 (R-A-SP-G-Q)
    Database link: P16949

  • Positive control
    • human breast carcinoma

Properties

Applications

Our Abpromise guarantee covers the use of ab47328 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/1000.
ICC/IF 1/100 - 1/500.
IHC-P 1/50 - 1/100.
ELISA 1/10000.

Target

  • Function
    Involved 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 specificity
    Ubiquitous. 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 similarities
    Belongs to the stathmin family.
    Contains 1 SLD (stathmin-like) domain.
  • Post-translational
    modifications
    Many 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.
  • Cellular localization
    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • C1orf215 antibody
    • Lag antibody
    • LAP 18 antibody
    • LAP18 antibody
    • Leukemia associated phosphoprotein p18 antibody
    • Leukemia-associated phosphoprotein p18 antibody
    • Metablastin antibody
    • Oncoprotein 18 antibody
    • OP 18 antibody
    • Op18 antibody
    • p18 antibody
    • p19 antibody
    • Phosphoprotein 19 antibody
    • Phosphoprotein p19 antibody
    • pp17 antibody
    • pp19 antibody
    • PR22 antibody
    • Pr22 protein antibody
    • Prosolin antibody
    • Protein Pr22 antibody
    • SMN antibody
    • Stathmin antibody
    • Stathmin1 antibody
    • STMN 1 antibody
    • Stmn1 antibody
    • STMN1_HUMAN antibody
    see all

Images

  • ab47328 staining human breast carcinoma tissue by IHC-P (left hand panel). The right hand panel shows staining in the presence of phospho-peptide.
  • ab47328 staining Stathmin1 (phospho S16) in Rat hippocampul neurons by Immunocytochemistry/ Immunofluorescence. Cultures of primary hippocampal neurons were prepared from embryonic day 18 rat embryos. The cells were transfected with GFP-shc (control shRNA vector) as control or GFP-sh2 Kidins220 / ARMS-specific shRNA, using serum-free medium with Lipofectamine. The cells were grown on coverslips and fixed for 5 min in 4% paraformaldehyde containing 4% sucrose in phosphate-buffered saline at 37 °C. Cells were then permeabilized with 0.2% Triton X-100 in phosphate-buffered saline during 5 min at room temperature. After blocking (5% bovine serum albumin in phosphate-buffered saline for 1 h), cells were incubated with the primary antibody. An Alexa Fluor®647 conjugated secondary antibody was used.

References

This product has been referenced in:
See all 5 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
Answer

Thank you for your reply. I am sorry that the antibodies did not work for you as stated on the datasheet. I have processed your request to receive refunds for all 3 of these antibodies. Your credit note number is ********. If there is anything else I can help you with, please let me know.

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Question

LOT NUMBER -- NOT SPECIFIED -- ORDER NUMBER 951895 DESCRIPTION OF THE PROBLEM Antibody is not specific for phosphorylated S37(38) on Stathmin-1 (STMN1). Antibody also recognises STMN1 without S37(38) phosphorylation. In other words, the antibody's recognition of STMN1 is not specific to the phosphorylated S37(38) residue. SAMPLE Whole cell lysate of colon carcinoma cell line HCT116. PRIMARY ANTIBODY ab47399 from Abcam Rabbit 1% milk in TBS-T (0.1% Tween, v/v) 1:1000 2hr at room temperature wash 3X5min DETECTION METHOD Pierce West Dura POSITIVE AND NEGATIVE CONTROLS USED Negative control: Phosphatase treated total cell lysate. ANTIBODY STORAGE CONDITIONS Upon receipt, antibodies were directly stored as 5µl aliquots at -20oC, without any freeze-thaw. For each experiment, a fresh aliquot was used. SAMPLE PREPARATION Freshly cultured cells were lysed in urea-based buffer (7M Urea, 2M Thiourea, 4% CHAPS, 10mM Tris) with DNase, RNase and protease inhibitor cocktail, without heating. AMOUNT OF PROTEIN LOADED A total of 20µg whole cell lysate was loaded per lane. ELECTROPHORESIS/GEL CONDITIONS Proteins were separated on 12.5% reducing SDS-PAGE. TRANSFER AND BLOCKING CONDITIONS Towbin’s transfer buffer was used, and gels were transferred at 100V for 1hr. Blocking was performed for 1 hr at room temperature with 5% non-fat milk in TBS + 0.1% Tween. SECONDARY ANTIBODY Goat anti-rabbit secondary antibody from Santa Cruz was used at 1:10000 dilution, for 1 hr at room temperature. Three 5 min washing steps were carried out. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Please refer to attached file.

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Answer

Thank you for contacting us. We have received your three western blot complaint questionnaires for ab47328, ab47398, and ab47399. These questionnaires refer to attachments with additional data, but unfortunately, we did not receive any attachments. It would be very helpful if you could please provide images of the blots for these antibodies, or tell me the molecular weights of the observed bands. Also, how long were the samples treated and what concentration of phosphatase was used when probing with ab47399? Milk contains endogenous phosphatases which may be causing the lack of specific signal observed with ab47328 and ab47398. If BSA has been giving poor results, you could try adding phosphatase inhibitors to your milk. These antibodies were validated using a reduced and denatured western blot protocol. Were the samples treated with SDS and a reducing agent such as BME or DTT prior to loading? It is possible that without being fully reduced and denatured (including boiling the samples for 5 minutes at 95C), the epitopes recognized by these antibodies may not exposed, which would prevent the antibody from binding effectively. I hope this helps, if not, please let me know and I will be happy to assist you further.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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