Huntingtin Recombinant Rabbit Monoclonal Antibody [JB89-34]
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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
概述
产品名称
Huntingtin Recombinant Rabbit Monoclonal Antibody [JB89-34]
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Synthetic peptide within N-terminal Human Huntingtin .
种属反应性
Human, Mouse, Rat
验证应用
WB, IHC-P, FC, IHC-Fr, IF-Cell, IF-Tissue
分子量
Predicted band size: 348 kDa
阳性对照
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.
偶联
unconjugated
克隆号
JB89-34
RRID
产品特性
形态
Liquid
浓度
1ug/ul
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
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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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1:50-1:200
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FC
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1:50-1:100
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IHC-Fr
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1:100
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IF-Cell
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1:100
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IF-Tissue
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1:50
靶点
功能
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.
背景文献
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).
序列相似性
Belongs to the huntingtin family.
组织特异性
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.
翻译后修饰
[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.
亚细胞定位
Cytoplasm. Nucleus.
别名
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
展开图片
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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. -
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Huntingtin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60, 1/50) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-Huntingtin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60, 1/50) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded mouse epididymis tissue using anti-Huntingtin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60, 1/50) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Huntingtin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60, 1/50) for 30 minutes 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.
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Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Huntingtin with Rabbit anti-Huntingtin antibody (ET7107-60).
The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET7107-60, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner. -
Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Huntingtin with Rabbit anti-Huntingtin antibody (ET7107-60).
The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET7107-60, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner. -
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. -
Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling Huntingtin with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/50 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET7107-60, green) at 1/50 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). -
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).
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