Huntingtin Recombinant Rabbit Monoclonal Antibody [JB89-34]
Catalog# ET7107-60
Huntingtin Recombinant Rabbit Monoclonal Antibody [JB89-34]
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WB
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IHC-P
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FC
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IHC-Fr
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IF-Cell
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IF-Tissue
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Human
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Mouse
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Rat
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Pig
Predicted species support after-sales service
Overview
Product Name
Huntingtin Recombinant Rabbit Monoclonal Antibody [JB89-34]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within N-terminal Human Huntingtin .
Species Reactivity
Human, Mouse, Rat (Predicted: Pig)
Predicted species support after-sales service
Validated Applications
WB, IHC-P, FC, IHC-Fr, IF-Cell, IF-Tissue
Molecular Weight
Predicted band size: 348 kDa
Positive Control
Neuro-2a cell lysate, PC-12 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, human colon carcinoma tissue, human breast tissue, mouse epididymis tissue, rat brain tissue, SH-SY5Y.
Conjugation
unconjugated
Clone Number
JB89-34
RRID
Product Features
Form
Liquid
Concentration
1ug/ul
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000-1:10,000
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IHC-P
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1:200-1:1,000
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FC
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1:50-1:100
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IHC-Fr
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1:200
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IF-Cell
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1:100
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IF-Tissue
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1:100-1:200
Target
Function
Huntingtin (Htt) is the protein coded for by the HTT gene, also known as the IT15 ("interesting transcript 15") gene. Mutated HTT is the cause of Huntington's disease (HD), and has been investigated for this role and also for its involvement in long-term memory storage. It is variable in its structure, as the many polymorphisms of the gene can lead to variable numbers of glutamine residues present in the protein. In its wild-type (normal form), it contains 6-35 glutamine residues. However, in individuals affected by Huntington's disease (an autosomal dominant genetic disorder), it contains more than 36 glutamine residues (highest reported repeat length is about 250). Its commonly used name is derived from this disease; previously, the IT15 label was commonly used. Within cells, huntingtin may or may not be involved in signaling, transporting materials, binding proteins and other structures, and protecting against apoptosis, a form of programmed cell death. The huntingtin protein is required for normal development before birth. It is expressed in many tissues in the body, with the highest levels of expression seen in the brain.
Background References
1. Cornett J et al. Polyglutamine expansion of huntingtin impairs its nuclear export. Nat Genet 37:198-204 (2005).
2. Sayer J A et al. Interaction of the nuclear matrix protein NAKAP with HypA and huntingtin: implications for nuclear toxicity in Huntington\'s disease pathogenesis. NeuroMolecular Med 7:297-310 (2005).
Sequence Similarity
Belongs to the huntingtin family.
Tissue Specificity
Expressed in the brain cortex (at protein level). Widely expressed with the highest level of expression in the brain (nerve fibers, varicosities, and nerve endings). In the brain, the regions where it can be mainly found are the cerebellar cortex, the neocortex, the striatum, and the hippocampal formation.
Post-translational Modification
[Huntingtin]: Cleaved by caspases downstream of the polyglutamine stretch. The resulting N-terminal fragments are cytotoxic and provokes apoptosis.; [Huntingtin]: Forms with expanded polyglutamine expansion are specifically ubiquitinated by SYVN1, which promotes their proteasomal degradation.; [Huntingtin]: Phosphorylation at Ser-1179 and Ser-1199 by CDK5 in response to DNA damage in nuclei of neurons protects neurons against polyglutamine expansion as well as DNA damage mediated toxicity.; [Huntingtin, myristoylated N-terminal fragment]: Myristoylated at Gly-551, following proteolytic cleavage at Asp-550.
Subcellular Location
Cytoplasm. Nucleus.
Synonyms
AI256365 antibody
C430023I11Rik antibody
HD antibody
HD protein antibody
HD_HUMAN antibody
HDH antibody
HTT antibody
Huntingtin antibody
HUNTINGTON CHOREA antibody
Huntington disease protein antibody
ExpandImages
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Western blot analysis of Huntingtin on different lysates with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/5,000 dilution.
Lane 1: Neuro-2a cell lysate (15 µg/Lane)
Lane 2: PC-12 cell lysate (15 µg/Lane)
Lane 3: Mouse brain tissue lysate (20 µg/Lane)
Lane 4: Rat brain tissue lysate (20 µg/Lane)
Predicted band size: 348 kDa
Observed band size: 348 kDa
Exposure time: 2 minutes 17 seconds;
3-8% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-60) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of Huntingtin on different lysates with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/10,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Huntingtin KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 348 kDa
Observed band size: 348 kDa
Exposure time: 100 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-60) at 1/10,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Application: IHC-Fr
Species: Mouse
Site: Cerebral cortex
Sample: Frozen section
Antibody concentration: 1/200
Antigen retrieval: Not required -
Application: IF-tissue
Species: Mouse
Site: Cerebral cortex
Sample: Paraffin-embedded section
Antibody concentration: 1/200 -
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunocytochemistry analysis of SH-SY5Y cells labeling Huntingtin with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of Huntingtin was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7107-60, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"