Huntingtin / Htt neoepitope 552 Antibody

Huntingtin / Htt neoepitope 552 Polyclonal Antibody for Western blot, IF, IHC, ELISA

>> See 11 other antibodies for HTT (Htt, Huntingtin, HD, IT15)
Synonyms:
Htt, Huntingtin, HD, IT15
Entrez Gene ID:
UniProt ID:
Details
Host / Isotype: Rabbit / IgG
Class: Polyclonal
Type: Antibody
Tested Species Reactivity: Human (Hu), Mouse (Ms)
Published Species Reactivity: Human (Hu)
Immunogen: Synthetic peptide conjugated to KLH via cysteine corresponding to residues SDPAMDLND (544-552) of Human HTT.
Ordering Information
Pierce Huntingtin / Htt neoepitope 552 Antibody
Product #
PA1-003
Size
100 µl
Price
$316.00
Purchase
Add Huntingtin / Htt neoepitope 552 Antibody to your cart.
Tested Applications Dilution *
Western Blot (WB) 1:500
Immunofluorescence (IF) 1:50 - 1:200
Immunohistochemistry (IHC) 1:50 : 1:200
ELISA (ELISA) 1:20 - 1:100
Published Applications Dilution
Western Blot (WB) See publications below
Immunofluorescence (IF) See publications below
Immunohistochemistry (IHC) See publications below
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Form Information
Form: Liquid
Concentration: 0.4mg/ml
Purification: Affinity chromatography
Storage Buffer: PBS with 1mg/ml BSA
Preservative: 0.05% sodium azide
Storage Conditions: -20°C
Product Specific Information
Neoepitope antibodies distinguish smaller cleaved fragments or processed forms of proteins versus the intact full-length or precursor by using a designed peptide purification process to maximize immunoreactivity to a specific cleavage site.

Human HTT caspase cleavage sites generate fragment-specific forms of the protein. Caspase-3/7 has been shown to generate cleavage sites at animo acids 513 and 552. Caspase-2 cleaves at amino acid 552 and caspase-6 at amino acid 586. Neo-specific antibody PA1-003 recognizes the 552 cleaved fragment without detecting the full-length form.
General Information
Huntingtin is a disease gene linked to Huntington's disease, a neurodegenerative disorder characterized by loss of striatal neurons. This is thought to be caused by an expanded, unstable trinucleotide repeat in the huntingtin gene, which translates as a polyglutamine repeat in the protein product. The huntingtin locus is large, spanning 180 kb and consisting of 67 exons. The huntingtin gene is widely expressed and is required for normal development. It is expressed as 2 alternatively polyadenylated forms displaying different relative abundance in various fetal and adult tissues. The larger transcript is approximately 13.7 kb and is expressed predominantly in adult and fetal brain whereas the smaller transcript of approximately 10.3 kb is more widely expressed. The genetic defect leading to Huntington's disease may not necessarily eliminate transcription, but may confer a new property on the mRNA or alter the function of the protein.
Product Images
  • Huntingtin / Htt neoepitope 552 Antibody (PA1-003) in WB
    PA1-003_WB_Image.jpg

    Western Blot with anti-Huntingtin  /  Htt neoepitope 552 Polyclonal Antibody (PA1-003) (PA1-003_WB_Image.jpg)

    Western Blot with anti-Huntingtin / Htt neoepitope 552 Polyclonal Antibody (PA1-003)

    Western blot analysis of endogenous HTT lysates with or without different caspase activity (Lanes 1-5) and overexpressed recombinant HTT fragment lysates (Lanes 6-9) was performed by loading 20 ug of lysate per well onto a 4-12% Bis-Tris polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 3% BSA/TBST for at least 1 hour. Membranes were then probed with a N-terminal pan-HTT antibody (top panel) or neoepitope-specific rabbit polyclonal antibody Product # PA1-003 (bottom panel) at a dilution of 1:500 overnight at 4°C on a rocking platform. Membranes were then washed in TBS-0.1%Tween 20 and probed with a goat anti-rabbit-HRP secondary antibody at 1:30,000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
  • Huntingtin / Htt neoepitope 552 Antibody (PA1-003) in IF
    Htt-neo-552_Antibody_PA1-003_Immunofluorescence_1.jpg

    Immunofluorescence with anti-Huntingtin  /  Htt neoepitope 552 Polyclonal Antibody (PA1-003) (Htt-neo-552_Antibody_PA1-003_Immunofluorescence_1.jpg)

    Immunofluorescence with anti-Huntingtin / Htt neoepitope 552 Polyclonal Antibody (PA1-003)

    Immunofluorescent analysis of Caspase cleaved Htt (green) in 293T cells transfected with an Htt23Q (left panel) and Htt148Q (right panel) stop constructs ending in amino acid 552. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 5% normal goat serum for 15 minutes at room temperature. Cells were probed with an Htt neo-epitope 552 polyclonal antibody (Product# PA1-003) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with goat-anti-rabbit secondary antibody at room temperature. Nuclei (blue) were stained with Hoechst dye.
  • Huntingtin / Htt neoepitope 552 Antibody (PA1-003) in IHC
    Htt-neo-552_Antibody_PA1-003_Immunohistochemistry_1.jpg

    Immunohistochemistry with anti-Huntingtin  /  Htt neoepitope 552 Polyclonal Antibody (PA1-003) (Htt-neo-552_Antibody_PA1-003_Immunohistochemistry_1.jpg)

    Immunohistochemistry with anti-Huntingtin / Htt neoepitope 552 Polyclonal Antibody (PA1-003)

    Immunohistochemistry was performed on tissues from transgenic HD mice of the YAC128 line. Cortex tissue samples were probed with an Htt neo-epitope 552 polyclonal antibody (Product #PA1-003) at a dilution of 1:50 overnight at 4°C in a humidified chamber. Tissues were washed extensively and endogenous peroxidase activity quenched for 30 minutes at room temperature. Detection was performed using a goat anti-rabbit HRP secondary antibody (green). Tissues were counterstained with nuclei stain (blue) and prepped for mounting.
  • Huntingtin / Htt neoepitope 552 Antibody (PA1-003) in ELISA
    Htt-neo-552_Antibody_PA1-003_ELISA_1.jpg

    ELISA with anti-Huntingtin  /  Htt neoepitope 552 Polyclonal Antibody (PA1-003) (Htt-neo-552_Antibody_PA1-003_ELISA_1.jpg)

    ELISA with anti-Huntingtin / Htt neoepitope 552 Polyclonal Antibody (PA1-003)

    Sandwich ELISA was performed with a monoclonal antibody to Htt and an Htt neo-epitope 552 polyclonal antibody (Product # PA1-003) to determine the antigen concentration of the Htt cleavage products. The curve represents a dose response for neo552 in 293T cells overexpressing the Htt construct.
Publications:
Western Blot
  Species / Dilution Summary
  Hu / Not Cited
PA1-003 was used in western blot, immunofluorescence, and immunohistochemistry to investigate the role of caspase-cleaved sites of mutant HTT in Huntington's Disease

J.Neurosci.. 2002;22(18):7862-7872.
"Caspase Cleavage of Mutant Huntingtin Precedes Neurodegeneration in Huntington's Disease."
Author(s): Wellington CL*, Ellerby LM*, Gutekunst CA, Rogers D, Warby S, Graham RK, Loubser O, van Raamsdonk J, Singaraja R, Yang YZ, Gafni J, Bredesen D, Hersch SM, Leavitt BR, Roy S, Nicholson DW, Hayden MR. *Co-first Authors
Number of Citations: 0
(See PubMed article )
  Hu / Not Cited
PA1-003 was used in western blot to validate the role of caspase activity in Huntington's Disease

Cell Death Differ.. 2004;11(4):424-438.
"Specific Caspase Interactions and Amplification Are Involved in Selective Neuronal Vulnerability in Huntington's Disease."
Author(s): Hermel E, Gafni J, Propp S, Leavitt B, Wellington CL, Hackam A, Logvinova A, Peel A, Chen S, Singaraja R, Karjewski S, Goldsmith PC, Ellerby HM, Salvesen GS, Hayden MR, Bredesen DE, Ellerby LM.
Number of Citations: 9
(See PubMed article )
  Hu / Not Cited
PA1-003 was used in western blot to validate the role of caspase inhibitors in Huntington's Disease

Chem.Biol.. 2012;17(11):1189-1200.
"Identification and Evaluation of Small Molecule Pan-Caspase Inhibitors in Huntington’s Disease Models. "
Author(s): Leyva MJ, DeGiacomo F, Kaltenbach LS, Holcomb J, Zhang N, Gafni J, Park H, Lo DC, Salvesen GS, Ellerby LM, Ellman JA.
Number of Citations: 1
(See PubMed article )
Immunofluorescence
  Species / Dilution Summary
  Hu / Not Cited
PA1-003 was used in western blot, immunofluorescence, and immunohistochemistry to investigate the role of caspase-cleaved sites of mutant HTT in Huntington's Disease

J.Neurosci.. 2002;22(18):7862-7872.
"Caspase Cleavage of Mutant Huntingtin Precedes Neurodegeneration in Huntington's Disease."
Author(s): Wellington CL*, Ellerby LM*, Gutekunst CA, Rogers D, Warby S, Graham RK, Loubser O, van Raamsdonk J, Singaraja R, Yang YZ, Gafni J, Bredesen D, Hersch SM, Leavitt BR, Roy S, Nicholson DW, Hayden MR. *Co-first Authors
Number of Citations: 0
(See PubMed article )
Immunohistochemistry
  Species / Dilution Summary
  Hu / Not Cited
PA1-003 was used in western blot, immunofluorescence, and immunohistochemistry to investigate the role of caspase-cleaved sites of mutant HTT in Huntington's Disease

J.Neurosci.. 2002;22(18):7862-7872.
"Caspase Cleavage of Mutant Huntingtin Precedes Neurodegeneration in Huntington's Disease."
Author(s): Wellington CL*, Ellerby LM*, Gutekunst CA, Rogers D, Warby S, Graham RK, Loubser O, van Raamsdonk J, Singaraja R, Yang YZ, Gafni J, Bredesen D, Hersch SM, Leavitt BR, Roy S, Nicholson DW, Hayden MR. *Co-first Authors
Number of Citations: 0
(See PubMed article )
(This product is for In Vitro experimental use only. Not for resale without express authorization.)
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