Western blot analysis of SMN on a HepG2 cell lysate (Human hepatocellular carcinoma; ATCC HB-8065) (left). Lane 1: 1:5000, lane 2: 1:10,000, lane 3: 1:20,000 dilution of the mouse anti-SMN antibody. Immunohistochemical staining of pyrimidal cells in a rat cortex, formalin-fixed paraffin-embedded tissue section, with citrate pre-treatment (magnification, 20X) (center). Immunofluorescent staining of differentiated SH-SY5Y cells (right). Cells were seeded in a 96 well, collagen coated imaging plate (Material # 353219) at ~ 5,000 cells per well. Cells were incubated with 50 mM ATRA (Sigma, R2625) for 5 days, followed by 50 ng/ml BDNF (Sigma, B3795) for 5 days. Differentiated cells were fixed and stained using the Triton X100 fix/perm protocol (see Recommended Assay Procedure; Bioimaging protocol link) and the anti-SMN antibody. The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen)(pseudo colored green). Cell nuclei were counter stained with Hoechst 33342 (pseudo colored blue). The image was taken on a BD Pathway™ 855 or 435 Bioimager System using a 20x objective and merged using the BD AttoVison ™ software This antibody also stained undifferentiated SH-SY5Y, SK-N-SH, C6, U87 and U373 cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).
Western blot analysis of SMN on a HepG2 cell lysate (Human hepatocellular carcinoma; ATCC HB-8065) (left). Lane 1: 1:5000, lane 2: 1:10,000, lane 3: 1:20,000 dilution of the mouse anti-SMN antibody. Immunohistochemical staining of pyrimidal cells in a rat cortex, formalin-fixed paraffin-embedded tissue section, with citrate pre-treatment (magnification, 20X) (center). Immunofluorescent staining of differentiated SH-SY5Y cells (right). Cells were seeded in a 96 well, collagen coated imaging plate (Material # 353219) at ~ 5,000 cells per well. Cells were incubated with 50 mM ATRA (Sigma, R2625) for 5 days, followed by 50 ng/ml BDNF (Sigma, B3795) for 5 days. Differentiated cells were fixed and stained using the Triton X100 fix/perm protocol (see Recommended Assay Procedure; Bioimaging protocol link) and the anti-SMN antibody. The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen)(pseudo colored green). Cell nuclei were counter stained with Hoechst 33342 (pseudo colored blue). The image was taken on a BD Pathway™ 855 or 435 Bioimager System using a 20x objective and merged using the BD AttoVison ™ software This antibody also stained undifferentiated SH-SY5Y, SK-N-SH, C6, U87 and U373 cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).
Product Details
BD Transduction Laboratories™
Survival Motor Neuron
Human (QC Testing), Mouse, Rat, Dog (Tested in Development)
Mouse IgG1
Human SMN aa. 14-174
Western blot (Routinely Tested), Bioimaging, Immunofluorescence, Immunohistochemistry-formalin (antigen retrieval required) (Tested During Development)
40 kDa
250 µg/ml
AB_397973
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at -20°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Triton is a trademark of the Dow Chemical Company.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
HRP Goat Anti-Mouse Ig RUO
Size
1 mL
Cat No.
554002
FITC Goat Anti-Mouse Ig RUO
Size
0.5 mg
Cat No.
554001
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Antibody Details
8/SMN
SMN (survival motor neuron) was discovered as a candidate gene, located in chromosome 5q13, for the fatal autosomal Spinal muscular atrophy (SMA) disorder. The SMN gene was missing or interrupted in a significant number of patients with SMA. The SMN protein is 294 amino acids and migrates with apparent molecular weight of 40 kDa. In addition to the cytoplasm, other studies localized SMN in dots of 0.1-1.0 µm within the nucleus. These novel nuclear structures were named "gems" and found associated to coiled bodies. It was also found that SMN interacts with the RGG box of hnRNP U and fibrillarin. Therefore, the biochemical function of SMN may be in the regulation of mRNA metabolism.
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Format Details
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
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Citations & References
Development References (5)
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Cifuentes-Diaz C, Frugier T, Tiziano FD. Deletion of murine SMN exon 7 directed to skeletal muscle leads to severe muscular dystrophy. J Cell Biol. 2001; 152(5):1107-1114. (Biology: Western blot). View Reference
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Claus P, Doring F, Gringel S. Differential intranuclear localization of fibroblast growth factor-2 isoforms and specific interaction with the survival of motoneuron protein. J Biol Chem. 2003; 278(1):479-485. (Biology: Immunoprecipitation, Western blot). View Reference
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Côté J, Boisvert FM, Boulanger MC, Bedford MT, Richard S. Sam68 RNA binding protein is an in vivo substrate for protein arginine N-methyltransferase 1. Mol Biol Cell. 2003; 14(1):274-287. (Biology: Immunoprecipitation, Western blot). View Reference
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Lefebvre S, Bürglen L, Reboullet S. Identification and characterization of a spinal muscular atrophy-determining gene. Cell. 1995; 80(1):155-165. (Biology). View Reference
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Wang IF, Reddy NM, Shen CK. Higher order arrangement of the eukaryotic nuclear bodies. Proc Natl Acad Sci U S A. 2002; 99(21):13583-13588. (Biology: Immunofluorescence, Western blot). View Reference
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Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
