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Monoclonal antibody against mouse Sox 17.

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Monoclonal antibody against mouse Sox 17. (SOX17 is a member of the SOX (SRY-related HMG-box) family of transcription factors .  The Sox family of proteins contains a high mobility group (HMG) motif that binds the DNA minor groove). Also known as VUR3.

 

Product Information

 

Product Number:                    S001

 

Size:                                        100ug

 

Concentration:                       0.5 mg/ml.

 

Storage Buffer:                       Phosphate buffered saline, pH 7.2 with 0.05% sodium azide.

 

Clone:                                     BC24-3.5CH 

 

Host:                                       Mouse

Immunogen:                            Recombinant mouse Sox 17 protein

 

Isotype:                                   Mouse IgG1, Kappa

 

Reactivity:                              The antibody was tested against mouse embryonic tissue and human, mouse and dog cell lines. Not tested in other species or in formalin fixed, paraffin embedded (FFPE) tissue. In most cells tested, the staining pattern observed is perinuclear.

                                                               

Applications:                           Indirect immunofluorescence, immunohistochemistry & western blotting. Not tested for use in immunoprecipitation. Suggested concentrations are as follows; Western blotting 2-3ug/ml, immunohistochemistry 3-5ug/ml. The BC24-3.5CH antibody may be used on tissue culture cells grown on chamber slides, cytospins and cryosections.  For staining, the cultured cells and cryosections should be fixed in 1-2% paraformaldehyde for 30 mins, permeabilized in 0.25% Triton X 100 in PBS for 30 mins and non-specific binding blocked with 1% BSA in PBS. The primary antibody may be used at 3-5ug/ml in 1%BSA in PBS. Please note that if used in situ for characterizing mouse embryos, some background staining may be observed, although this will not detract from the specific staining (see immunoperoxidase staining below). The reflects the presence of mouse serum immunoglobulin in the sections which are detected by the anti-mouse Ig secondary antibody. This may be avoided by using directly labeled 3.5CH without the need for a labeled anti mouse Ig secondary.  This problem should not be observed for embryos of other species. 

 

Preparation and Storage:      The monoclonal antibody is purified from tissue culture supernatant using Protein G sephasore columns. Store undiluted at 4° C or aliquot and store at -20C for long term storage. Repeated freeze thaw cycles should be avoided.

 

Western blotting-:                   Use the BC24-3.5CH antibody at 2-3 ug/ml in Tris buffered Saline with 0.05% Tween 20 and 5% non-fat dry milk (Blotto) or similar diluents.

Western blotting with 3.5CH anti-SOX-17 antibody (BC24-3.5CH) used at 2 ug/ml in Blotto on Sox-17 MDCK cell lysate loaded at 20ug/lane.

 

Secondary antibody: Peroxidase conjugated rabbit anti-mouse Ig used at 1/20,000 dilution in Blotto. Developed by enhanced chemiluminescence.

Predicted molecular size. Although the predicted molecular size of Sox 17 is approximately 45 kDa, a higher band of above 50kd is observed.  It is thought that the high proline content of Sox 17 reduces its electrophoretic mobility thus giving it an apparent higher molecular weight of above 50kD rather than the predicted molecular size of about 45kDa.

Suggested positive control cell lines:  TF1-1a (ATCC CRL-2451) hematopoietic cell line and MDCK (NBL-2) (ATCC CCL-34) cell lines are suggested as positive control cell lines. The antibody reacts with a band of approximate molecular size 50kDa. 

Although Sox 17 was first characterized in embryonal development, it has also been shown to be expressed in some cancers. Our unpublished characterization of the developed antibodies show that the protein is expressed in various types of cancer cell lines and as shown in the previous page, the expression of Sox 17 may be modulated by drugs such as Aphidicolin and Nocadazole that block cells in Go or G2M stage of the cell cycle respectively. That Sox 17 is expressed in the G2M stage of the cell cycle is interesting and warrants further study.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: The product contains 0.05% sodium azide. Sodium azide produces highly toxic hydrazoic acid under acidic conditions. It is therefore recommended to dilute azide compounds in running water before discarding to prevent the accumulation of potentially explosive deposits in the plumbing.

 

Background.                                                                                                        

SOX17 is a member of the SOX (SRY-related HMG-box) family of transcription factors that play important roles in embryonic development and in the determination of the cell fate. The Sox family of proteins contains a high mobility group (HMG) motif that binds the DNA minor groove and are often used as markers to assess the differentiation of specific cell lineages. Whereas the association of Oct 4 with Sox 2 imparts pluripotent cell fate, the association of Oct 4 with Sox 17 imparts an  endodermal cell fate. The expression pattern of Sox 17 is complex.  It is expressed in progenitor cells derived from two different germ layers and activates the transcription of key regulator genes for vasculogenesis, hematopoiesis, and erythrocyte differentiation. Sox 17 plays a key role in endoderm formation, cardiac myogenesis, kidney and urinary development and differentiation of oligodendrocytes. The expression of Sox 17 has been used as a marker of lineage or disease in various studies. The overexpression of Sox 17 is thought to promote tumor angiogenesis and vascular abnormalities and therefore tumor progression. Sox 17 expression is associated with gastric cancer progression, whereas SOX2 and SOX17 expression patterns are thought to be useful for distinguishing between seminoma and embryonal carcinoma. Sox17 was shown to be specifically expressed in fetal and neonatal but not adult HSCs and may therefore be a useful marker for distinguishing between embryonic and adult stem cells.

 

References

 

Yang H., Lee S., Lee S., Kim K., Yang Y., Kim JH., Adams RH., Wells JM., Morrison SJ., Koh GY., Kim I.Sox17 promotes tumor angiogenesis and destabilizes tumor vessels in mice. J Clin Invest. 2013;123(1):418-31.

 

Takayama K., Inamura M,. Kawabata K., Tashiro K., Katayama K., Sakurai F., Hayakawa T., Furue MK., Mizuguchi H. Efficient and directive generation of two distinct endoderm lineages from human ESCs and iPSCs by differentiation stage-specific SOX17 transduction. PLoS One. 2011;6(7):e21780.

Séguin CA., Draper JS., Nagy A., Rossant J. Establishment of endoderm progenitors by SOX transcription factor expression in human embryonic stem cells. Cell Stem Cell. 2008;3(2):182-95.

 

Aksoy I., Jauch R., Chen J., Dyla M., Divakar U., Bogu GK., Teo R., Leng Ng CK., Herath W., Lili S., Hutchins AP., Robson P., Kolatkar PR., Stanton LW. Oct4 switches partnering from Sox2 to Sox17 to reinterpret the enhancer code and specify endoderm.EMBO J. 2013;32(7):938-53.

 

Nakajima-Takagi Y., Osawa M., Oshima M, Takagi H., Miyagi S., Endoh M., Endo TA., Takayama N., Eto K., Toyoda T., Koseki H., Nakauchi H., Iwama A. Role of SOX17 in hematopoietic development from human embryonic stem cells. Blood. 2013;121(3):447-58.

Ye YW., Wu JH., Wang CM., Zhou Y., Du CY., Zheng BQ., Cao X., Zhou XY., Sun MH., Shi YQ. Sox17 regulates proliferation and cell cycle during gastric cancer progression. Cancer Lett. 2011; 307(2):124-31.

 

Fukamachi H., Shimada S., Ito K., Ito Y., Yuasa Y. CD133 is a marker of gland-forming cells in gastric tumors and Sox17 is involved in its regulation. Cancer Sci. 2011; 102(7):1313-21.

 

 Wang P., Rodriguez RT., Wang J., Ghodasara A., Kim SK. Targeting SOX17 in human embryonic stem cells creates unique strategies for isolating and analyzing developing endoderm. Cell Stem Cell. 2011;8(3):335-46.

 

Niakan KK., Ji H., Maehr R., Vokes SA., Rodolfa KT., Sherwood RI., Yamaki M., Dimos JT., Chen AE., Melton DA., McMahon AP., Eggan K. Sox17 promotes differentiation in mouse embryonic stem cells by directly regulating extraembryonic gene expression and indirectly antagonizing self-renewal. Genes Dev. 2010;24(3):312-26.

 

Stefanovic S., Abboud N., Désilets S., Nury D., Cowan C., Pucéat M. Interplay of Oct4 with Sox2 and Sox17: a molecular switch from stem cell pluripotency to specifying a cardiac fate. J Cell Biol. 2009;186(5):665-73.

 

Du YC., Oshima H., Oguma K., Kitamura T., Itadani H., Fujimura T., Piao YS., Yoshimoto T., Minamoto T., Kotani H., Taketo MM., Oshima M. Induction and down-regulation of Sox17 and its possible roles during the course of gastrointestinal tumorigenesis. Gastroenterology. 2009;137(4):1346-57.

Nonaka D. Differential expression of SOX2 and SOX17 in testicular germ cell tumors. Am J Clin Pathol. 2009;131(5):731-6.

Kim I., Saunders TL., Morrison SJ. Sox17 dependence distinguishes the transcriptional regulation of fetal from adult hematopoietic stem cells. Cell. 2007 Aug 10;130(3):470-83.

 

Sohn J., Natale J., Chew LJ., Belachew S., Cheng Y., Aguirre A., Lytle J., Nait-Oumesmar B., Kerninon C., Kanai-Azuma M., Kanai Y., Gallo V. Identification of Sox17 as a transcription factor that regulates oligodendrocyte development. J Neurosci. 2006;26(38):9722-35.

 

Kanai Y., Kanai-Azuma M., Noce T., Saido TC., Shiroishi T., Hayashi Y., Yazaki K. Identification of two Sox17 messenger RNA isoforms, with and without the high mobility group box region, and their differential expression in mouse spermatogenesis. J Cell Biol. 1996;133(3):667-81.


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