MAB1286
Anti-Nuclear Spliceosomes Antibody, clone 780-3
clone 780-3, Chemicon®, from mouse
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
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biological source
mouse
antibody form
purified from hybridoma cell culture
antibody product type
primary antibodies
clone
780-3, monoclonal
species reactivity
human
manufacturer/tradename
Chemicon®
technique(s)
immunofluorescence: suitable
immunohistochemistry: suitable
western blot: suitable
isotype
IgM
shipped in
wet ice
target post-translational modification
unmodified
Specificity
Stains nuclear spliceosomes. Using indirect immunofluorescence, produces a speckled nuclear staining patter. Immunoelectron microscopy has shown that the antibody recognizes a discrete area of the nucleus identified as interchromatin granules. Immunoblots 105 kD molecular weight protein from human cells. Immunoprecipitates spliceosome and low-levels of snRNPs 1,2,5 (using 0.5 M salt).
Application
Detect Nuclear Spliceosomes with Anti-Nuclear Spliceosomes Antibody, clone 780-3 (Mouse Monoclonal Antibody), that has been shown to work in WB, IHC & IF.
Indirect immunofluorescence: 1:20-1:30. Use 30-100 μl per well/slide.
Optimal working dilutions must be determined by the end user.
In addition to the lymphnoid cultures, normal human epithelial cells can be screened by indirect immunofluorescence microscopy for positive reactions with the hybridoma supernatants. Since the human epithelial cells grow as monolayer cultures, they are plated directly onto the printed microscope slides after trypsinization and allowed to attach and grow overnight at 37°C in complete medium. The following day, the slides are briefly rinsed in PBS to remove the medium and the cells are fixed as described above. In general, the slides are not allowed to air dry either during or after the fixation procedure in order to maintain the cellular integrity and antigenicity of intracellular molecules.
For photographic analysis, viable cell preparations obtained from Ficol-hypaque gradient separations are cytocentrifuged directly onto slides at 1250 rpm for 10 minutes. This procedure flattens the lymphoid cells and greatly improves the visibility of intranuclear and cytoplasmic antigens. Slides prepared in this manner are fixed in the same way directly after cytocentrifugation.
In order to screen the hybridoma supernatants by indirect immunofluorescence, 30-100 μL of each supernatant (optimize for each individual assay) are pipetted on a well of the printed microscope slides using a different tip for each supernatant. After 60 minutes of incubation at 37°C in a humidified chamber, the slides are rinsed 3 times with PBS at room temperature, and again incubated for 30 minutes at 37°C with 20 μL of a 1:20 dilution of fluorescein-conjugated goat anti-mouse IgG antibody (Millipore AP124F). The slides are then prepared rinsed 3 times with PBS, counterstained with Evans Blue for 5 minutes at room temperature using a freshly prepared solution containing 50 μL of a 1% stock solution of Evans Blue in 80 mL of PBS, rinsed a final time in PBS, and coverslipped using a 1:1 solution of glycerol: PBS. The slides are then examined by epifluorescence microscopy. Since many of the monoclonal antibodies produced a rapidly diminishing fluorescent reaction, exposure times optimally are less than five seconds.
Optimal working dilutions must be determined by the end user.
In addition to the lymphnoid cultures, normal human epithelial cells can be screened by indirect immunofluorescence microscopy for positive reactions with the hybridoma supernatants. Since the human epithelial cells grow as monolayer cultures, they are plated directly onto the printed microscope slides after trypsinization and allowed to attach and grow overnight at 37°C in complete medium. The following day, the slides are briefly rinsed in PBS to remove the medium and the cells are fixed as described above. In general, the slides are not allowed to air dry either during or after the fixation procedure in order to maintain the cellular integrity and antigenicity of intracellular molecules.
For photographic analysis, viable cell preparations obtained from Ficol-hypaque gradient separations are cytocentrifuged directly onto slides at 1250 rpm for 10 minutes. This procedure flattens the lymphoid cells and greatly improves the visibility of intranuclear and cytoplasmic antigens. Slides prepared in this manner are fixed in the same way directly after cytocentrifugation.
In order to screen the hybridoma supernatants by indirect immunofluorescence, 30-100 μL of each supernatant (optimize for each individual assay) are pipetted on a well of the printed microscope slides using a different tip for each supernatant. After 60 minutes of incubation at 37°C in a humidified chamber, the slides are rinsed 3 times with PBS at room temperature, and again incubated for 30 minutes at 37°C with 20 μL of a 1:20 dilution of fluorescein-conjugated goat anti-mouse IgG antibody (Millipore AP124F). The slides are then prepared rinsed 3 times with PBS, counterstained with Evans Blue for 5 minutes at room temperature using a freshly prepared solution containing 50 μL of a 1% stock solution of Evans Blue in 80 mL of PBS, rinsed a final time in PBS, and coverslipped using a 1:1 solution of glycerol: PBS. The slides are then examined by epifluorescence microscopy. Since many of the monoclonal antibodies produced a rapidly diminishing fluorescent reaction, exposure times optimally are less than five seconds.
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors
Transcription Factors
Physical form
Format: Purified
Hybridoma supernatant, concentrated by ammonium sulfate. Buffer: PBS with 0.1% sodium azide.
Storage and Stability
Maintain at -20°C in undiluted aliquots for up to 12 months. Avoid repeated freeze/thaw cycles.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Storage Class
10 - Combustible liquids
wgk_germany
WGK 2
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificates of Analysis (COA)
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Akira Inoue et al.
Biology of the cell, 100(9), 523-535 (2008-03-05)
The RNA-binding protein S1-1, also called RBM10 (RNA-binding motif 10), is a paralogue of putative tumour suppressor RBM5 and has been correlated with cancer proliferation and apoptosis. In the present study, we have investigated the cell biology of S1-1. In
The MUC1 extracellular domain subunit is found in nuclear speckles and associates with spliceosomes.
Priyadarsini Kumar et al.
PloS one, 7(8), e42712-e42712 (2012-08-21)
MUC1 is a large transmembrane glycoprotein and oncogene expressed by epithelial cells and overexpressed and underglycosylated in cancer cells. The MUC1 cytoplasmic subunit (MUC1-C) can translocate to the nucleus and regulate gene expression. It is frequently assumed that the MUC1
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