Q: Could you recommond the Human specific SMN antibody? It will only recognize human SMN, without cross reaction with mouse smn.

A: 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.

Q: I was wondering if the Alexa 647 conjugated anti-SMN antibody (NB100-1936AF647) can be used in Western Blot, and if so, the recommended working concentration is 2ug/ml?

A: Our SMN antibody with catalogue number NB100-1936AF647 is covered by our 100% quality guarantee for the Western blotting application. The recommended starting dilution for the unconjugated antibody, NB100-1936, is 2ug/ml. With an unconjugated antibody however the signal will be amplified through the use of a conjugated secondary for detection. Since NB100-1936AF647 is directly conjugated, and no secondary antibody is required for detection, it is possible that you might need to use a higher concentration of the antibody to obtain a satisfactory signal. We do always recommend that our customers optimise their staining conditions to suit their particular sample type, and you might wish to try a number of different dilutions when using this antibody initially.

SMN Antibody (2B1) [Alexa Fluor® 647]

Novus Biologicals | Catalog # NB100-1936AF647

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Product Specifications
Scientific Data
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Background
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Primate, Xenopus, Zebrafish

Cited:

Fish - Danio rerio (Zebrafish)

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Immunocytochemistry/ Immunofluorescence

Label

Alexa Fluor 647 (Excitation = 650 nm, Emission = 668 nm)

Antibody Source

Monoclonal Mouse IgG₁ Clone # 2B1

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Product Specifications

Immunogen

Purified recombinant His6-tagged human SMN protein. [Swiss-Prot# Q16637]

Reactivity Notes

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

Localization

Cytoplasm. Nucleus > Gem, localized in subnuclear structures next to coiled bodies, called Gemini of Cajal bodies (Gems).

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Scientific Data Images for SMN Antibody (2B1) [Alexa Fluor® 647]

SMN-Antibody-2B1-Alexa-Fluor-R-647-Immunocytochemistry-Immunofluorescence-NB100-1936AF647-img0003.jpg

SMN (2B1) in U-2 OS Human Cell Line -

SMN (2B1) was detected in immersion fixed U-2 OS human osteosarcoma cell line using Mouse anti-SMN (2B1) Protein G Purified Monoclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NB100-1936AF647) (light blue) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using a 100X objective and digitally deconvolved.

Detection of SMN (2B1) in A431 Human Cell Line by Flow Cytometry.

An intracellular stain was performed on A431 human skin carcinoma cell line using Mouse anti- SMN (2B1) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NB100-1936AF647, blue histogram) or matched control antibody (orange histogram) at 2.5 µg/mL for 30 minutes at RT.

Immunocytochemistry/ Immunofluorescence: SMN Antibody (2B1) [Alexa Fluor® 647] [NB100-1936AF647] -

Immunocytochemistry/ Immunofluorescence: SMN Antibody (2B1) [Alexa Fluor® 647] [NB100-1936AF647] - Etv5b activates the smn promoter leading to increased Smn levels in motoneurons.(a–d) Whole-mount in situ hybridization of etv5b expression in 24 hpf embryos. Expression pattern in the entire embryo (a), in magnified lateral (b) & dorsal (c) views as well as in a transverse section (d) of the trunk. In the latter case, yellow arrowhead denote signal in the motoneuron region. Scale bars: 100 μm (a) & 50 μm (b–d). nc, notochord. (e) RT-PCR in control (C - only psmn:mCherry) versus etv5b overexpressing (OE - etv5b mRNA & psmn:mCherry) embryos at early gastrula stage. (f,g) scIF of control (f) & etv5b MO (g) cells from pmnx1:eGFP embryos. H2B, GFP & Smn signals are shown in Z-projected confocal sections. GFP+ cells marked by white rectangles are magnified on the right. Scale bars: 10 μm for low & 5 μm for high magnification. (h) Relative Smn signal in control & in etv5b MO cells from one representative experiment. Exact values are (mean ± SD): 0.99 ± 0.25 (control) & 0.7 ± 0.14 (etvb5 MO), p = 0.003 with Wilcoxon Sum Rank Test. (i) Average relative Smn levels in control & in etv5b MO motoneurons. The exact value of decrease is 0.75 ± 0.04 (mean ± SD). N = 3 experiments, n = number of analyzed cells. Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep27470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: SMN Antibody (2B1) [Alexa Fluor® 647] [NB100-1936AF647] -

Immunocytochemistry/ Immunofluorescence: SMN Antibody (2B1) [Alexa Fluor® 647] [NB100-1936AF647] - Motoneurons exhibit elevated Smn levels.(a–c) Single-cell immunofluorescent (scIF) experiments. 24 hpf pmnx1:eGFP embryos were dissociated & immunostained for Smn ((a) see also Methods). DIC image of cells following 3-hour (b) & 24-hour (c) incubation are shown with GFP in green. Note that the GFP signal in the cell body is oversaturated so that the weaker signal in the axon becomes visible. In (c) DNA is in blue & the motoneuron marker Znp1 (synaptotagmin) is in red; the axon growth cone is magnified in the corner of the images. Scale bars are 100 μm (a), 10 μm (low magnification in (b,c)) & 2 μm (high magnification in (c)). (d) scIF on pmnx1:eGFP embryos. Histone 2B (H2B), GFP & Smn signals are shown in Z-projected confocal sections. Cells marked by white rectangles are magnified on the right. Scale bars: 10 μm for low & 5 μm for high magnification. (e) To account for potential variability in the immunostaining, Smn levels were quantified relative to H2B. Diamonds denote GFP negative (GFP−) & GFP positive (GFP+) cells from one representative experiment. Blue bars indicate mean ± SD with significance values of *p < 0.05 & **p < 0.01. Exact values are (mean ± SD) 0.51 ± 0.14 (GFP−) & 0.87 ± 0.17 (GFP+), p = 0.001 with Wilcoxon Sum Rank Test. For more details, see Materials & Methods & Supplementary Table S1. (f) Average increase of Smn levels in motoneurons versus control cells. The exact value of enrichment is (mean ± SD): 1.67 ± 0.14. N = 3 experiments, n = number of analyzed cells. Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep27470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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