SMN Antibody (2B1)

Immunohistochemistry: SMN Antibody (2B1) [NB100-1936] - CBs lose their colocalization with SMN nuclear foci in diabetic mice. Normal and pathological distribution of SMN proteins in sensory neurons. SMN proteins were found throughout the cytoplasm and as nuclear foci in control sensory neurons. SMN nuclear foci localized within CBs (coilin) in controls (upper panels, white arrows), whereas in the diabetic nucleus the numerous CBs were present but lost their colocalization with SMN nuclear foci (lower panels, yellow arrows). Arrowheads indicate sensory neurons magnified in the insets. Scale bars: 20 um, 10 um in insets. Image collected and cropped by CiteAb from the following publication (//dmm.biologists.org/lookup/doi/10.1242/dmm.028225), licensed under a CC-BY license.

Western Blot: SMN Antibody (2B1) [NB100-1936] - Analysis of SMN expression in 1) HeLa, 2) NTera2, 3) HepG2, 4) MCF7, 5) NIH 3T3, 6) PC12, and 7) COS7 whole cell lysates.

Immunocytochemistry/Immunofluorescence: SMN Antibody (2B1) [NB100-1936] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- NB100-1936 at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunocytochemistry/Immunofluorescence: SMN Antibody (2B1) [NB100-1936] - scIF of control (f) and etv5b MO (g) cells from pmnx1:eGFP embryos. H2B, GFP and Smn signals are shown in Z-projected confocal sections. GFP+ cells marked by white rectangles are magnified on the right. Scale bars: 10 um for low and 5 um for high magnification. Image collected and cropped by Citeab from the following publication (Transcriptional enhancement of Smn levels in motoneurons is crucial for proper axon morphology in zebrafish. Sci Rep (2016) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: SMN Antibody (2B1) [NB100-1936] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with SMN Antibody [2B1] conjugated to Alexa Fluor 488 (NB100-1936AF488) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: SMN Antibody (2B1) [NB100-1936] - An intracellular stain was performed on Neuro2a cells with SMN Antibody (2B1) NB100-1936 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Immunocytochemistry/Immunofluorescence: SMN Antibody (2B1) [NB100-1936] - The SMN antibody was tested at a 1:250 dilution in HeLa cells against DyLight 488 (Green). Actin nuclei were counterstained against Phalloidin 568 (Red) and DAPI (Blue), respectively.

Immunohistochemistry: SMN Antibody (2B1) [NB100-1936] - Analysis of SMN on mouse brain using NB100-1936.

Flow Cytometry: SMN Antibody (2B1) [NB100-1936] - An intracellular stain was performed on Ntera2 cells with SMN Antibody (2B1) NB100-1936 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

• Reactivity: Hu, Mu, Rt, Pm, Xp, Ze
• Applications: WB, ELISA, Flow, ICC/IF, IHC, IP
• Clone: 2B1
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

SMN Antibody (2B1) Summary

• Immunogen: Purified recombinant His6-tagged human SMN protein. [Swiss-Prot# Q16637]
• Localization: Cytoplasm. Nucleus > Gem, localized in subnuclear structures next to coiled bodies, called Gemini of Cajal bodies (Gems).
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: SMN1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • ELISA reported in scientific literature (PMID 23973875)
  • Flow Cytometry reported in scientific literature (PMID 30102724)
  • Immunocytochemistry/Immunofluorescence 1:250
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:200
  • Immunoprecipitation 1:10-1:500
  • Western Blot 2 ug/ml
• Application Notes: This SMN Antibody (2B1) is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry on paraffin-embedded sections, Flow Cytometry, Immunoprecipitation, and Western blot where a band can be seen at ~ 35 kDa.
• Control:

  HeLa Whole Cell Lysate

Reactivity Notes

Zebrafish reactivity reported in scientific literature (PMID: 27273160).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine and 0.15M NaCl
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for SMN Antibody (2B1)

• Gemin-1
• Kugelberg-Welander disease)
• SMNC
• survival motor neuron protein
• survival of motor neuron 1, telomeric

Background

Spinal muscular atrophy (SMA) is a common fatal autosomal recessive disorder characterized by the degeneration of motor neurons and muscular atrophy. A gene called Survival of Motor Neurons (SMN) has been identified as the determining gene in SMA and is deleted or mutated in greater than 98% of SMA patients. The SMN protein localizes to both the cytoplasm and the nucleus. In the nucleus, SMN has been found to localize in a nuclear structure termed gems for Gemini of coiled bodies, which contain high levels of small ribonucleoproteins (snRNPs). SMN forms complexes with proteins including SIP1, Gemin 2-7 and several others known to be involved in the biogenesis of snRNPs. SMN antibodies may serve as a useful research tool to study SMN complexes and gems, and their role in pre-RNA processing and RNA metabolism.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for SMN Antibody (NB100-1936)(23)

Publications using NB100-1936

• Wu SH, Lu IC, Yang SM et al. Spinal Irisin Gene Delivery Attenuates Burn Injury-Induced Muscle Atrophy by Promoting Axonal Myelination and Innervation of Neuromuscular Junctions International journal of molecular sciences 2022-12-14 [PMID: 36555538] (WB, Rat)
• Stewart LM, Gerner L, Rettel M Et al. CaMKK2 facilitates Golgi-associated vesicle trafficking to sustain cancer cell proliferation Cell death & disease 2021-11-01 [PMID: 34725334]
• Nishibe M, Toyoda H, Hiraga SI Et al. Synaptic and Genetic Bases of Impaired Motor Learning Associated with Modified Experience-Dependent Cortical Plasticity in Heterozygous Reeler Mutants Cerebral cortex (New York, N.Y. : 1991) 2021-07-31 [PMID: 34339488] (IF/IHC, Mouse)
• Otsuki N, Arakawa R, Kaneko K et al. A new biomarker candidate for spinal muscular atrophy: Identification of a peripheral blood cell population capable of monitoring the level of survival motor neuron protein PLoS ONE. 2018-08-13 [PMID: 30102724] (FLOW, Human)
  
  Details:
  Citation using the Alexa Fluor 488 form of this antibody.
• Chen HY, Lin YN, Chen WC et al. Urethral proteomic analysis in ovariectomized mice administered 17b-oestradiol replacement therapy. J Obstet Gynaecol. 2017-03-28 [PMID: 28350532] (Mouse)
• Kobayashi M, Chandrasekhar A, Cheng C et al. Diabetic polyneuropathy, sensory neurons, nuclear structure and spliceosome alterations: a role for CWC22. Dis Model Mech. 2017-03-01 [PMID: 28250049]
• Spiro Z, Koh A, Tay S et al. Transcriptional enhancement of Smn levels in motoneurons is crucial for proper axon morphology in zebrafish. Sci Rep 2016-06-09 [PMID: 27273160] (ICC/IF, Zebrafish)
  
  Details:
  This publication used the AF647 conjugated form of this antibody (NB100-1936AF647)
• Kassim SH, Jordan J, Schreiter J et al. Systematic identification of novel SLE related autoantibodies responsible for type I IFN production in human plasmacytoid dendritic cells. Cell Immunol. 2013-08-06 [PMID: 23973875] (ELISA, Human)
• Yong J, Golembe TJ, Battle DJ, Pellizzoni L, Dreyfuss G. snRNAs contain specific SMN-binding domains that are essential for snRNP assembly. Mol Cell Biol;24(7):2747-56. 2004-04-01 [PMID: 15024064] (IP, WB, Xenopus, Human)
• Nelson PT, Hatzigeorgiou AG, Mourelatos Z. miRNP:mRNA association in polyribosomes in a human neuronal cell line. RNA. 2004-03-01 [PMID: 14970384] (WB, Human)
• Massenet S, Pellizzoni L, Paushkin S, Mattaj IW, Dreyfuss G. The SMN complex is associated with snRNPs throughout their cytoplasmic assembly pathway. Mol Cell Biol;22(18):6533-41. 2002-09-01 [PMID: 12192051] (IP, Human)
• Paushkin S, Gubitz AK, Massenet S, Dreyfuss G. The SMN complex, an assemblyosome of ribonucleoproteins. Curr Opin Cell Biol;14(3):305-12. 2002-06-01 [PMID: 12067652] (IP, Human)
• Pellizzoni L, Baccon J, Rappsilber J, Mann M, Dreyfuss G. Purification of native survival of motor neurons complexes and identification of Gemin6 as a novel component. J Biol Chem;277(9):7540-5. 2002-03-01 [PMID: 11748230] (IP, Human)
• Gubitz AK, Mourelatos Z, Abel L, Rappsilber J, Mann M, Dreyfuss G. Gemin5, a novel WD repeat protein component of the SMN complex that binds Sm proteins. J Biol Chem;277(7):5631-6. 2002-02-15 [PMID: 11714716] (WB, IP, Human)
• Friesen WJ, Massenet S, Paushkin S, Wyce A, Dreyfuss G. SMN, the product of the spinal muscular atrophy gene, binds preferentially to dimethylarginine-containing protein targets. Mol Cell;7(5):1111-7. 2001-05-01 [PMID: 11389857] (IP, WB, Human)
• Kerr DA, Nery JP, Traystman RJ, Chau BN, Hardwick JM. Survival motor neuron protein modulates neuron-specific apoptosis. Proc Natl Acad Sci U S A;97(24):13312-7. 2000-11-21 [PMID: 11078511] (WB, Mouse)
• Charroux B, Pellizzoni L, Perkinson RA, Yong J, Shevchenko A, Mann M, Dreyfuss G. Gemin4. A novel component of the SMN complex that is found in both gems and nucleoli. J Cell Biol;148(6):1177-86. 2000-03-20 [PMID: 10725331] (IP, ICC/IF, WB, Human)
• Giesemann et al. A role for polyproline motifs in the spinal muscular atrophy protein SMN. Profilins bind to and colocalize with smn in nuclear gems. J Biol Chem;274(53):37908-14. 1999-12-31 [PMID: 10608857] (ICC/IF, Human)
• Pellizzoni L, Kataoka N, Charroux B, Dreyfuss G. A novel function for SMN, the spinal muscular atrophy disease gene product, in pre-mRNA splicing. Cell;95(5):615-24. 1998-11-25 [PMID: 9845364] (ICC/IF, Human)
• La Bella V, Cisterni C, Salaun D, Pettmann B. Survival motor neuron (SMN) protein in rat is expressed as different molecular forms and is developmentally regulated. Eur J Neurosci;10(9):2913-23. 1998-09-01 [PMID: 9758161] (WB, Human, Rat)
• Fischer U, Liu Q, Dreyfuss G. The SMN-SIP1 complex has an essential role in spliceosomal snRNP biogenesis. Cell;90(6):1023-9. 1997-09-19 [PMID: 9323130] (WB, IP, Xenopus)
• Liu Q, Fischer U, Wang F, Dreyfuss G. The spinal muscular atrophy disease gene product, SMN, and its associated protein SIP1 are in a complex with spliceosomal snRNP proteins. Cell;90(6):1013-21. 1997-09-19 [PMID: 9323129] (WB, ICC/IF, IP, Human)
• Liu Q, Dreyfuss G. A novel nuclear structure containing the survival of motor neurons protein. EMBO J;15(14):3555-65. 1996-07-15 [PMID: 8670859] (IP, WB, ICC/IF, Human, Mouse, Xenopus)

FAQs for SMN Antibody (NB100-1936). (Showing 1 - 1 of 1 FAQs).

1. Could you recommond the Human specific SMN antibody? It will only recognize human SMN, without cross reaction with mouse smn.
   • We do not have an SMN antibody that has been validated NOT to work in mouse samples. When an antibody is found to not react with a certain species, we mark the data sheet with a (-) next to the species' name. Unfortunately, the SMN antibodies that do not list mouse have not been tested in mouse, and may react.

Bioinformatics

• Gene Symbol: SMN1
• Entrez:
  • 6606(Human)
  • 20595(Mouse)
  • 64301(Rat)
• Uniprot:
  • Q16637(Human)
  • P97801(Mouse)
  • O35876(Rat)