Name: Anti-Huntingtin antibody [EPR5526]
Brand: RabMAb
SKU: ab109115
Rating: 5 (4 reviews)

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Western blot - Anti-Huntingtin antibody [EPR5526] (ab109115)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR5526] to Huntingtin
Suitable for: Flow Cyt (Intra), IHC-FoFr, ICC/IF, WB, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 APC Carrier Free PE

製品名

Anti-Huntingtin antibody [EPR5526]
Huntingtin 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR5526] to Huntingtin
由来種

Rabbit
アプリケーション

適用あり: Flow Cyt (Intra), IHC-FoFr, ICC/IF, WB, IHC-Pmore details
種交差性

交差種: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: SH-SY5Y, HeLa, HAP1, PC-12 and Neuro-2a whole cell lysates; Mouse and rat brain lysates. IHC-P: Human cerebral cortex and astrocytoma tissue; Mouse and rat testis tissue. ICC/IF: Neuro-2a and SH-SY5Y cell lines. Flow Cyt (intra): SH-SY5Y cell line. IHC-Fr: Mouse and rat cerebrum.
特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
バッファー

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR5526
アイソタイプ

IgG
研究分野

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human HTT (Huntingtin) knockout HeLa cell line (ab265976)
KO cell lysates
- Human HTT (Huntingtin) knockout HeLa cell lysate (ab256946)
Recombinant Protein
- Recombinant Human Huntingtin protein (Tagged) (ab112300)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab109115の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
Flow Cyt (Intra)		1/250. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-FoFr		1/100. Perform Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF		1/1000.
WB	(3)	1/5000. Detects a band of approximately 348 kDa (predicted molecular weight: 348 kDa).
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

特記事項
Flow Cyt (Intra) 1/250. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-FoFr 1/100. Perform Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF 1/1000.
WB 1/5000. Detects a band of approximately 348 kDa (predicted molecular weight: 348 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

機能

May play a role in microtubule-mediated transport or vesicle function.
組織特異性

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.
関連疾患

Defects in HTT are the cause of Huntington disease (HD) [MIM:143100]. HD is an autosomal dominant neurodegenerative disorder characterized by involuntary movements (chorea), general motor impairment, psychiatric disorders and dementia. Onset of the disease occurs usually in the third or fourth decade of life and symptoms progressively worsen leading to death in 10 to 20 years. Onset and clinical course depend on the degree of poly-Gln repeat expansion, longer expansions resulting in earlier onset and more severe clinical manifestations. HD affects 1 in 10,000 individuals of European origin. Neuropathology of Huntington disease displays a distinctive pattern with loss of neurons, especially in the caudate and putamen (striatum).
配列類似性

Belongs to the huntingtin family.
Contains 10 HEAT repeats.
ドメイン

The N-terminal Gln-rich and Pro-rich domain has great conformational flexibility and is likely to exist in a fluctuating equilibrium of alpha-helical, random coil, and extended conformations.
翻訳後修飾

Cleaved by apopain downstream of the polyglutamine stretch. The resulting N-terminal fragment is cytotoxic and provokes apoptosis.
Forms with expanded polyglutamine expansion are specifically ubiquitinated by SYVN1, which promotes their proteasomal degradation.
細胞内局在

Cytoplasm. Nucleus. The mutant Huntingtin protein colocalizes with AKAP8L in the nuclear matrix of Huntington's disease neurons.
Target information above from: UniProt accession P42858 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 3064 Human
- Entrez Gene: 15194 Mouse
- Entrez Gene: 29424 Rat
- Omim: 143100 Human
- Omim: 613004 Human
- SwissProt: P42858 Human
- SwissProt: P42859 Mouse
- SwissProt: P51111 Rat
- Unigene: 518450 Human
- Unigene: 209071 Mouse
- Unigene: 482929 Mouse
- Unigene: 11193 Rat
see all
別名
- 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
- Huntington's disease protein homolog antibody
- IT 15 antibody
- IT15 antibody
- OTTMUSP00000026909 antibody
- ZHD antibody
see all

Western blot - Anti-Huntingtin antibody [EPR5526] (ab109115)

All lanes : Anti-Huntingtin antibody [EPR5526] (ab109115) at 1/10000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : HTT knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 348 kDa
Observed band size: 348 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab109115 observed at 348 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab109115 was shown to react with Huntingtin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265976 (knockout cell lysate ab256946) was used. Wild-type HeLa and HTT knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109115 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Huntingtin antibody [EPR5526] (ab109115)

Immunohistochemistry (Frozen) analysis of mouse cerebellum tissue sections labeling Huntingtin with purified ab109115 at 1/100 (13.4 µg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. Sections were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Negative control: PBS instead of the primary antibody. DAPI (blue) was used as nuclear counterstain. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) was performed.
Immunocytochemistry/ Immunofluorescence - Anti-Huntingtin antibody [EPR5526] (ab109115)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized

Neuro-2a (Mouse neuroblastoma cells) cells labeling Huntingtin with purified ab109115 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear and cytoplasmic staining on Neuro-2a cell line.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows:
1. ab191472 at 1/1000 dilution followed by ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor^® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Western blot - Anti-Huntingtin antibody [EPR5526] (ab109115)

All lanes : Anti-Huntingtin antibody [EPR5526] (ab109115) at 1/10000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : HTT knockout HAP1 whole cell lysate
Lane 3 : SH-SY5Y whole cell lysate
Lane 4 : HeLa whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 348 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab109115 observed at 348 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab109115 was shown to specifically recognize HTT in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when HTT knockout samples were exmined. Wild-type and HTT knockout samples were subjected to SDS-PAGE. Unpurified ab109115 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10,000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] (ab109115)

Immunohistochemical analysis of paraffin-embedded Human astrocytoma labeling Huntingtin with purified ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Nuclear staining on cancer cells of astrocytoma.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Huntingtin antibody [EPR5526] (ab109115)

Immunohistochemistry (Frozen) analysis of mouse cerebrum tissue sections labeling Huntingtin with purified ab109115 at 1/100 (13.4 µg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. Sections were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Negative control: PBS instead of the primary antibody. DAPI (blue) was used as nuclear counterstain. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) was performed.
Western blot - Anti-Huntingtin antibody [EPR5526] (ab109115)

All lanes : Anti-Huntingtin antibody [EPR5526] (ab109115) at 1/5000 dilution (purified)

Lane 1 : SH-SY5Y (Human neuroblastoma from bone marrow cells) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : Neuro-2a (Mouse neuroblastoma cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 348 kDa
Observed band size: 348 kDa

Exposure time: 1 second

Blocking and Diluting buffer and concentration: 5% NFDM /TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] (ab109115)

Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Huntingtin with purified ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Nuclear staining on neuron of human cerebral cortex was observed.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Huntingtin antibody [EPR5526] (ab109115)

All lanes : Anti-Huntingtin antibody [EPR5526] (ab109115) at 1/50000 dilution (purified)

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 348 kDa
Observed band size: 348 kDa

Exposure time: 30 seconds

Blocking and Diluting buffer and concentration: 5% NFDM /TBST
Flow Cytometry (Intracellular) - Anti-Huntingtin antibody [EPR5526] (ab109115)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling Huntingtin with purifiedab109115 at 1/250 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Huntingtin antibody [EPR5526] (ab109115)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized

SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling Huntingtin with purified ab109115 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear and cytoplasmic staining on SH-SY5Y cell line.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows:
1. ab191472 at 1/1000 dilution followed by ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor^® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] (ab109115)

Immunohistochemical analysis of paraffin-embedded Mouse testis labeling Huntingtin with purified ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Cytoplasmic staining on spermatogenic cells of mouse testis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] (ab109115)

Immunohistochemical analysis of paraffin-embedded Rat testis labeling Huntingtin with purified ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Weak cytoplasmic staining on spermatogenic cells of rat testis.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-Huntingtin antibody [EPR5526] (ab109115)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

ab109115 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab109115 は 20 報の論文で使用されています。

Vigont VA et al. STIM2 Mediates Excessive Store-Operated Calcium Entry in Patient-Specific iPSC-Derived Neurons Modeling a Juvenile Form of Huntington's Disease. Front Cell Dev Biol 9:625231 (2021). PubMed: 33604336
Wang L et al. miR-146a Overexpression in Oral Squamous Cell Carcinoma Potentiates Cancer Cell Migration and Invasion Possibly via Targeting HTT. Front Oncol 10:585976 (2020). PubMed: 33282738
Schedin-Weiss S et al. Proteomics Time-Course Study of App Knock-In Mice Reveals Novel Presymptomatic Aß42-Induced Pathways to Alzheimer's Disease Pathology. J Alzheimers Dis 75:321-335 (2020). PubMed: 32280097
Axenhus M et al. Huntingtin Levels are Elevated in Hippocampal Post-Mortem Samples of Alzheimer's Disease Brain. Curr Alzheimer Res 17:858-867 (2020). PubMed: 33272184
Vernizzi L et al. Glutamine Synthetase 1 Increases Autophagy Lysosomal Degradation of Mutant Huntingtin Aggregates in Neurons, Ameliorating Motility in a Drosophila Model for Huntington's Disease. Cells 9:N/A (2020). PubMed: 31941072

View all Publications for this product

レビューと Q&A

Show All レビュー Q&A

レビューを送る質問を送る

1-5 of 5 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (neurons)

Gel Running Conditions

Reduced Denaturing

Loading amount

15 µg

Specification

neurons

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Nov 26 2021

Application

Immunoprecipitation

Sample

Mouse Tissue lysate - whole (Brain)

Total protein in input

1400 µg

Immuno-precipitation step

Other - Dynabeads

Specification

Brain

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Svitlana Yablonska

Verified customer

投稿 Apr 19 2017

Application

Western blot

Sample

Escherichia coli Purified protein (Purified protein)

Gel Running Conditions

Reduced Denaturing (10% gel)

Loading amount

2 µg

Treatment

Protease cleavage 3hrs

Specification

Purified protein

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Ya Zhou

Verified customer

投稿 Jun 28 2016

Application

Western blot

Sample

Bacteria Cell lysate - whole cell (BL21)

Gel Running Conditions

Non-reduced Denaturing (10% gel)

Loading amount

10 µg

Treatment

IPTG 1mM 16h

Specification

BL21

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jun 27 2016

Question

Inquiry: Hello, I was hoping you would be able to tell me which amino acids your anti-huntingtin antibody 109115 was specifically raised against? Your data sheet just says 'N-terminal.' Thank you,

Abcam community

Verified customer

Asked on Sep 24 2012

Answer

Thank you for contacting us.

The immunogen is from N-term from amino acid XXXX of P42858.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
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Abcam Scientific Support

Answered on Sep 24 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

組織特異性

関連疾患

配列類似性

ドメイン

翻訳後修飾

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

Datasheet download

参考文献 (20)

レビューと Q&A

Western blot abreview for Anti-Huntingtin antibody [EPR5526]

Immunoprecipitation abreview for Anti-Huntingtin antibody [EPR5526]

Western blot abreview for Anti-Huntingtin antibody [EPR5526]

Western blot abreview for Anti-Huntingtin antibody [EPR5526]