MAB5374
Anti-Huntingtin Protein Antibody, clone mEM48
culture supernatant, clone mEM48, Chemicon®
Synonym(s):
Huntingtin
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
mouse
Quality Level
antibody form
culture supernatant
antibody product type
primary antibodies
clone
mEM48, monoclonal
species reactivity
human
species reactivity (predicted by homology)
mouse, rat
manufacturer/tradename
Chemicon®
technique(s)
immunocytochemistry: suitable
immunohistochemistry: suitable
western blot: suitable
isotype
IgG
NCBI accession no.
UniProt accession no.
target post-translational modification
unmodified
Gene Information
human ... HTT(3064) , SLC6A4(6532)
General description
Huntington disease (HD) is a hereditary, progressive, neurodegenerative ailment characterized by personality changes, motor impairment and subcortical dementia. The molecular basis of the disease involves the expansion of the trinucleotide CAG, coding for polyglutamine in the first exon of a chromosome four gene (4p16.3), which normally produces a widely expressed 3136 a.a. (~350 kDa) protein huntingtin with unclear function. The protein is found in the perinuclear region along with microtubules, and in the centrosomal region along with gamma-tubulin. Huntingtin is necessary for neuronal survival and is involved in synaptic vesicle trafficking, microtubule binding and may also have a role in apoptosis. In the HD condition, neuronal cells with the mutant form of huntingtin possess intranuclear aggregations of the N-terminal fragment, causing damaging inclusions in perinuclear locations and striatal neuron cell death. Wild-type huntington and anti-huntingtin reduce aggregation and cellular toxicity of the mutant huntingtin form in mammalian cell models of HD. Huntingtin is known to interact with GAPDH, HAP-1, SP1 and TAFII130.
Specificity
Reacts with human huntingtin protein (both native and recombinant protein). MAB5374 reacts with mutant huntingtin in patients and in transgenic animals that express different numbers of repeats (from 82 to 150 glutamines). Thus, it should recognize different forms of mutant huntingtin.
Immunogen
GST fusion protein from the first 256 amino acids from human huntingtin with the deletion of the polyglutamine tract.
Application
Detect Huntingtin Protein using this Anti-Huntingtin Protein Antibody, clone mEM48 validated for use in IC, IH & WB.
Immunohistochemistry:
1:50-1:100 of a previous lot using ABC on 4% paraformaldehyde fixed tissue. Suggested dilution buffer is PBS containing 3% BSA. The antibody works on paraffin embedded tissue sections.
Suggested dilution buffer is PBS containing 3% BSA. The antibody works on paraffin embedded tissue sections. Yu, Z et al (2002) Hum. Mole. Genetics 11(8):905-914. (http://hmg.oxfordjournals.org/cgi/content/full/11/8/905) for good IHC methods and photos of mEM48 on rodent tissues with human transgenic material.
Immunocytochemistry:
light 4% PFA fixation followed by 0.1% triton X-100 incubation prior to blocking is suggested.
A previous lot of this antibody was used in IC.
Western blot:
1:50-1:500 using ECL depending on the level of mutant protein. Suggested dilution buffer is PBS containing 3% BSA or PBS containing 5% non-fat milk.
Nuclear fraction preparations enhance signals; monomeric protein ~80kDa; aggregates are common which can be >200kDa in size.
Optimal working dilutions must be determined by the end user.
1:50-1:100 of a previous lot using ABC on 4% paraformaldehyde fixed tissue. Suggested dilution buffer is PBS containing 3% BSA. The antibody works on paraffin embedded tissue sections.
Suggested dilution buffer is PBS containing 3% BSA. The antibody works on paraffin embedded tissue sections. Yu, Z et al (2002) Hum. Mole. Genetics 11(8):905-914. (http://hmg.oxfordjournals.org/cgi/content/full/11/8/905) for good IHC methods and photos of mEM48 on rodent tissues with human transgenic material.
Immunocytochemistry:
light 4% PFA fixation followed by 0.1% triton X-100 incubation prior to blocking is suggested.
A previous lot of this antibody was used in IC.
Western blot:
1:50-1:500 using ECL depending on the level of mutant protein. Suggested dilution buffer is PBS containing 3% BSA or PBS containing 5% non-fat milk.
Nuclear fraction preparations enhance signals; monomeric protein ~80kDa; aggregates are common which can be >200kDa in size.
Optimal working dilutions must be determined by the end user.
Research Category
Neuroscience
Neuroscience
Research Sub Category
Neurodegenerative Diseases
Neurodegenerative Diseases
Quality
Routinely evaluated by Western Blot on HEK293 lysates.
Western Blot Analysis:
1:1000 dilution of this lot detected Huntingtin Protein on 10 μg of HEK293 lysates.
Western Blot Analysis:
1:1000 dilution of this lot detected Huntingtin Protein on 10 μg of HEK293 lysates.
Target description
over 200 kDa
Physical form
Culture Supernatant mouse monoclonal IgG containing no preservative.
Unpurified
Storage and Stability
Stable for 6 months at -20ºC in undiluted aliquots from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Analysis Note
Control
Normal human cerebral cortex lysate, mouse brain cortex samples from HD or wild type mice
HEK293 lysates.
Normal human cerebral cortex lysate, mouse brain cortex samples from HD or wild type mice
HEK293 lysates.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
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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recommended
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 2
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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An engineered viral protease exhibiting substrate specificity for a polyglutamine stretch prevents polyglutamine-induced neuronal cell death.
Sellamuthu, S; Shin, BH; Han, HE; Park, SM; Oh, HJ; Rho, SH; Lee, YJ; Park, WJ
Testing null
Neuroprotective effects of inositol 1,4,5-trisphosphate receptor C-terminal fragment in a Huntington's disease mouse model.
Tang, TS; Guo, C; Wang, H; Chen, X; Bezprozvanny, I
The Journal of Neuroscience null
Inhibition of the striatal specific phosphodiesterase PDE10A ameliorates striatal and cortical pathology in R6/2 mouse model of Huntington's disease.
Giampa, C; Laurenti, D; Anzilotti, S; Bernardi, G; Menniti, FS; Fusco, FR
Testing null
Tomomi Imamura et al.
Scientific reports, 6, 33861-33861 (2016-09-23)
We identified drug seeds for treating Huntington's disease (HD) by combining in vitro single molecule fluorescence spectroscopy, in silico molecular docking simulations, and in vivo fly and mouse HD models to screen for inhibitors of abnormal interactions between mutant Htt
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