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Purified Mouse Anti-S100B
Purified Mouse Anti-S100B
Western blot analysis of S100B on a mouse cerebellum lysate (left).  Lane 1: 1:5000, lane 2: 1:10,000, lane 3: 1:20,000 dilution of the mouse anti-S100B antibody. Immunofluorescent staining of differentiated SH-SY5Y cells (Human neuroblastoma; ATCC CRL-2266) (right).  Cells were seeded in collagen coated 96-well imaging microplates (Material # 353219) at ~ 5,000 cells per well.  Cells were incubated with 50 µM all-trans-retinoic acid (ATRA) (Sigma-Aldrich, cat.no. R2625) for 5 days, followed by 50 ng/ml BDNF (Sigma-Aldrich, cat.no. B3795)  for 5 days.  Differentiated cells were fixed and stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and the mouse anti-S100B  antibody.  The second step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen).  The image was taken on a BD Pathway™ 855 or 435 Bioimager system using a 20x objective.  This antibody also stained undifferentiated SK-N-SH (Human neuroblastoma; ATCC HTB-11), C6 (Rat glioma; ATCC CCL-107), U-87 MG (Human glioblastoma cells; ATCC HTB-14) and U-373 cells (Human glioblastoma cells; ATCC HTB-17; discontinued, investigators may refer to: http://www.atcc.org/MisidentifiedCellLines/tabid/683/Default.aspx ) using both the Triton-X 100 and methanol fix/perm protocols (see Recommended Assay Procedure).
Western blot analysis of S100B on a mouse cerebellum lysate (left).  Lane 1: 1:5000, lane 2: 1:10,000, lane 3: 1:20,000 dilution of the mouse anti-S100B antibody. Immunofluorescent staining of differentiated SH-SY5Y cells (Human neuroblastoma; ATCC CRL-2266) (right).  Cells were seeded in collagen coated 96-well imaging microplates (Material # 353219) at ~ 5,000 cells per well.  Cells were incubated with 50 µM all-trans-retinoic acid (ATRA) (Sigma-Aldrich, cat.no. R2625) for 5 days, followed by 50 ng/ml BDNF (Sigma-Aldrich, cat.no. B3795)  for 5 days.  Differentiated cells were fixed and stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and the mouse anti-S100B  antibody.  The second step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen).  The image was taken on a BD Pathway™ 855 or 435 Bioimager system using a 20x objective.  This antibody also stained undifferentiated SK-N-SH (Human neuroblastoma; ATCC HTB-11), C6 (Rat glioma; ATCC CCL-107), U-87 MG (Human glioblastoma cells; ATCC HTB-14) and U-373 cells (Human glioblastoma cells; ATCC HTB-17; discontinued, investigators may refer to: http://www.atcc.org/MisidentifiedCellLines/tabid/683/Default.aspx ) using both the Triton-X 100 and methanol fix/perm protocols (see Recommended Assay Procedure).
商品详情
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BD Transduction Laboratories™
Mouse (QC Testing), Human, Rat (Tested in Development)
Mouse IgG1
Mouse S100B aa. 1-92
Bioimaging, Western blot (Routinely Tested), Immunofluorescence (Tested During Development)
8 kDa
250 µg/ml
AB_647296
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


推荐的实验流程

Western blot:  Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml

Bioimaging:

Methanol Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample.

Triton-X 100 Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.

商品通知

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
612376 Rev. 1
抗体详情
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19/S100B

The S100 multigene family consists of a number of small dimeric molecules around 100 residues in length. The original members of this family, S100A and S100B, were initially identified in bovine brain. Other S100 proteins have also been identified, such as S100P from human placenta and S100L from bovine lung. The S100 family consists of at least 10 members which show a cell-type-specific expression pattern. Most S100 proteins are believed to be involved in Ca2+ signaling pathways. S100P consists of 95 amino acids and shares about 50% sequence identity with S100A and S100B, while S100L has been shown to possess 43-47% homology to S100A and S100B. S100B is expressed in the nervous system by glial cells, such as astrocyte, oligodendroglial, ependymal, and microglial cells. The dimeric form of S100B has been implicated in neuronal survival, neurite extension, and astrocyte growth. In addition, S100B expression is induced by stimulations that activate p53, and S100B activity may be involved in PKC-dependent p53 nuclear translocation in early G1 phase. Thus, S100B plays a role in calcium signaling pathways that regulate cell differentiation and cell cycle progression.

612376 Rev. 1
格式详情
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
612376 Rev.1
报价单和参考
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研发参考 (2)

  1. Jiang H, Shah S, Hilt DC. Organization, sequence, and expression of the murine S100 beta gene. Transcriptional regulation by cell type-specific cis-acting regulatory elements. J Biol Chem. 1993; 268(27):20502-20511. (Biology). 查看参考
  2. Scotto C, Delphin C, Deloulme JC, Baudier J. Concerted regulation of wild-type p53 nuclear accumulation and activation by S100B and calcium-dependent protein kinase C. Mol Cell Biol. 1999; 19(10):7168-7180. (Biology). 查看参考
612376 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.