Antibody Panel to Huntingtin and Huntingtin Interacting Protein (HIP1, HIP2, HIP14) - FocusOn 011

Huntingtin is a 350 kDa protein of 3144 amino acids that is altered in Huntington's disease (HD). The expanded trinucleotide CAG repeat of the HD gene encodes an abnormally expanded polyglutamine stretch in the N-terminus of the Huntingtin protein. The normal function of Huntingtin still remains uncertain but it has been suggested that it performs a necessary housekeeping function. The abnormal form of Huntingtin aggregates in vitro and forms neuronal intranuclear and cytoplasmic inclusions in HD (Huntington’s Disease ) patients. The expanded polyglutamine repeats formed by this mutation of the Huntingtin gene have been proposed to cause neuronal degeneration in HD through abnormal interactions with other proteins containing short polyglutamine tracts such as the CREB binding protein (CBP), a protein that promotes cell survival and is a major mediator of survival signals in mature neurons.

Fig. 1: AP07921PU-N HAP1 antibody staining of Formalin-Fixed Paraffin-Embedded Human Adrenal tissue.
Fig. 1: AP07921PU-N HAP1 antibody staining of Formalin-Fixed Paraffin-Embedded Human Adrenal tissue.
Fig. 2: Total protein extract of normal human cerebral cortex separated as a strip on a 3-12.5% gradient SDS-PAGE gel and Western blotted. The blot was probed with SM1660.
Fig. 2: Total protein extract of normal human cerebral cortex separated as a strip on a 3-12.5% gradient SDS-PAGE gel and Western blotted. The blot was probed with SM1660.
Fig. 3: Western blot analysis of Huntington Disease protein in different lymphoblast cell line extracts (Lanes 1-4) (CAG repeats ~65 and ~20) using AM20340PU-N Huntingtin antibody (Panel A) and an antibody from another source (Panel B).
Fig. 3: Western blot analysis of Huntington Disease protein in different lymphoblast cell line extracts (Lanes 1-4) (CAG repeats ~65 and ~20) using AM20340PU-N Huntingtin antibody (Panel A) and an antibody from another source (Panel B).

It could be shown, that the IGF-1/Akt pathway is neuroprotective in Huntington's Disease and involves Huntingtin phosphorylation by Akt.

Huntingtin Interacting Protein 1 (HIP-1)

HIP-1 is a reportedly proapoptotic, cargo-specific adaptor protein that may be involved in the pathogenesis of Huntington Disease. As well as playing a role in HD, it is likely to be involved in the recruitment of clathrin coats to lipid membranes and it may also factor in tumorigenesis by allowing the survival of precancerous and cancerous cells.

HIP2 (Ubiquitin-conjugating enzyme E2 K, UBE2K, LIG)

HIP2 is a ubiquitin conjugating enzyme that binds selectively to a large region at the N terminus of huntingtin. HIP2-driven covalent modification of huntingtin with ubiquitin marks it for selective degradation via the proteasomal pathway. HIP2 may mediate foam cell formation by the suppression of apoptosis of lipid bearing macrophages through ubiquitination and subsequent degradation of p53.

Fig. 4: Human Brain, Cortex: Formalin-Fixed, Paraffin-Embedded (FFPE).
Fig. 4: Human Brain, Cortex: Formalin-Fixed, Paraffin-Embedded (FFPE). Cat.-No. AP23819PU-N.
 

HIP9 (AP2A2, ADTAA, CLAPA1)

HIP9 is the alpha subunit of the adaptor protein complex 2 (AP-2) which participates in membrane traffic pathways. AP-2 plays a key role in clathrin-dependent endocytosis in which cargo proteins are ferried into vesicles surrounded by clathrin (clathrin-coated vesicles, CCVs) designated for fusion with the early endosome. HIP9 (AP-2 alpha subunit) serves its connecting role by orienting AP-2 on the membrane (binding accessible polyphosphoinositide-containing lipids) while also recognizing and binding to the endocytosis signal motif [ED]-X-X-X-L-[LI] of endocytic transmembrane accessory proteins via its C-terminal cytosolic tail.

Fig. 5: Western blot using Affinity Purified anti-AP2A antibody shows detection of a band just below 100 kDa corresponding to Human AP2A1 in a various preparations. Lane 1 - HeLa nuclear extract, Lane 2 - HeLa, Lane 3 - 293, Lane 4 - A431 and Lane 5 - Jurkat whole cell lysates. In lanes 6-10 the antibody was preincubated with 1 µg/ml of the immunizing peptide which effectively blocks the specific reactivity of this antibody with AP2A. Approximately 20 µg of each lysate was run on a SDS-PAGE and transferred onto nitrocellulose followed by reaction with a 1:500 dilution of anti-AP2A antibody. Detection occurred using a 1:5,000 dilution of HRP-labeled Rabbit anti-Goat IgG for 1 hour at room temperature. A chemi-luminescence system was used for signal detection (Roche) using a 60-sec exposure time.
Fig. 5: Western blot using Affinity Purified anti-AP2A antibody shows detection of a band just below 100 kDa corresponding to Human AP2A1 in a various preparations. Lane 1 - HeLa nuclear extract, Lane 2 - HeLa, Lane 3 - 293, Lane 4 - A431 and Lane 5 - Jurkat whole cell lysates. In lanes 6-10 the antibody was preincubated with 1 µg/ml of the immunizing peptide which effectively blocks the specific reactivity of this antibody with AP2A. Approximately 20 µg of each lysate was run on a SDS-PAGE and transferred onto nitrocellulose followed by reaction with a 1:500 dilution of anti-AP2A antibody. Detection occurred using a 1:5,000 dilution of HRP-labeled Rabbit anti-Goat IgG for 1 hour at room temperature. A chemi-luminescence system was used for signal detection (Roche) using a 60-sec exposure time. Cat.-No. AP09207PU-N
 

Optineurin (OPT, FIP2, GLC1E, HIP7, HYPL, NRP)

Optineurin plays a neuroprotective role in the eye and optic nerve, with mutations in the gene associated with pathogenesis of gluacoma. Optineurin is implicated in signal transduction of the tumor necrosis factor (TNF) pathway, shifting the equilibrium toward induction of apoptotic cell death. A role for optineurin is also indicated in membrane traffic regulation and cellular morphogenesis, working proximate to cell membranes in concert with Rab8 and Huntingtin.

Fig. 6: Human Brain, Cerebellum (formalin-fixed, paraffin-embedded) stained with OPTN antibody AP22648PU-N at 2.5 µg/ml followed by biotinylated goat anti-rabbit IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.
Fig. 6: Human Brain, Cerebellum (formalin-fixed, paraffin-embedded) stained with OPTN antibody AP22648PU-N at 2.5 µg/ml followed by biotinylated goat anti-rabbit IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.
 

Huntingtin-associated protein-1 (HAP1, HAP2, HLP1)

Huntingtin-associated protein-1 is highly expressed in brain and demonstrated to mediate the neuropathology of HD. Huntingtin-associated protein-1 interacts with huntingtin, with two cytoskeletal proteins (dynactin and pericentriolar autoantigen protein 1), and with a hepatocyte growth factor-regulated tyrosine kinase substrate. The interactions with cytoskeletal proteins and a kinase substrate indicate a role for Huntingtin-associated protein-1 in vesicular trafficking or organelle transport.

A novel Huntingtin interacting protein is HIP14. Its interaction with Huntingtin is inversely correlated to the polyglutamine length in Huntingtin. HIP14 protein, which is enriched in the brain, has been reported to show partial co-localization with Huntingtin in the striatum. It is found in a subset of neurons affected in HD.

Antibody Tools for Detection of Huntingtin and HIP1, HIP2, HIP14

Acris Antibodies offers a comprehensive panel of monoclonal and polyclonal antibodies to Huntingtin and Huntingtin interacting Proteins. These antibodies can be used in Western blotting, some of them are also suitable for immunohistochemical applications or immunoprecipitation.

References

Wilkinson FL et al (1999) Localization of rabbit huntingtin using a new panel of monoclonal antibodies. Molecular Brain Research 69: 10 – 20

Mills, I.G., et al. Huntington interacting protein 1 modulates the transcriptional activity of nuclear hormone receptors. J. Cell Biol. 170(2): 191-200, 2005.

Singaraja Roshni R. ; Hadano Shinji; Metzler Martina ; Givan Scott ; Wellington Cheryl L.; Warby Simo ; Yanai Anat ; Gutekunst Claire-Anne ; Leavaitt Blair R. ; Hong YI ; Fichter Keith ; Gan Lu ; McCutcheon Krista ; Chopra Vikramjit; Michel Jennifer ; Hersch Steven M.; Ikeda J.-E; Hayden Michael R.: HIP14, a novel ankyrin domain-containing protein, links huntingtin to intracellular trafficking and endocytosis. Hum. mol. genet. ISSN 0964-6906 . 2002, 11(23): 2815-2828

Available antibody panel to Huntingtin and Huntingtin Interacting Protein-1, -2, -7, -9, 12, -13, -14 and HAP1

Huntingtin, Huntingtin interacting Protein-1 (HIP1), Huntingtin interacting Protein-2 (HIP2), Huntingtin interacting Protein-7 (HIP7 or Optineurin), Huntingtin interacting Protein-9 antibody (HIP9 or AP2A2), Huntingtin interacting Protein-12 antibody (HIP12), Huntingtin interacting Protein-13 (HIP13 or FICD) Huntingtin interacting Protein-14 (HIP14 or ZDHHC17), Huntingtin associated Protein (HAP1) and Huntingtin pSer421 antibody (Huntingtin phospho-specific).

13 Item(s)

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

AP2A2 / HIP9 Control Peptide

AP2A2 / HIP9
50 µg / €190.00
  Acris Antibodies GmbH

HAP1 antibody

Brain, cortex: Formalin-Fixed Paraffin-Embedded (FFPE) Rabbit IgG Aff - Purified Hu, Ms, Rt P, WB
50 µg / €470.00
  Acris Antibodies GmbH

HAP1 Control Peptide

HAP1
50 µg / €160.00
  Acris Antibodies GmbH

HIP2 / UBE2K (144-157) antibody

Human Brain, Cortex: Formalin-Fixed, Paraffin-Embedded (FFPE) Goat Aff - Purified Bear, Bov, Bt, Can, Chk, Eq, Fi, Gt, Hst, Hu, Mky, Ms, Op, Por, Prim, Rb, Rt, Tr, Xen E, P, WB
50 µg / €470.00
  Acris Antibodies GmbH

HIP12 / HIP1R (N-term) antibody

Western blot analysis of HIP12 / HIP1R Antibody (N-term) Cat.-No AP52048PU-N in Ramos cell line lysates (35ug/lane). This demonstrates the HIP1R antibody detected the HIP1R protein (arrow). Rabbit Ig Aff - Purified Hu F, P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

Huntingtin antibody

Human Brain, Cerebellum: Formalin-Fixed, Paraffin-Embedded (FFPE) Mouse 3F1 Purified Hu E, P, WB
50 µg / €530.00
  Acris Antibodies GmbH

Huntingtin control peptide

Huntingtin Purified
50 µg / €190.00
  Acris Antibodies GmbH

Huntingtin control peptide

Huntingtin Purified
50 µg / €190.00
  Acris Antibodies GmbH

Huntingtin pSer421 antibody

Figure 2. Immunohistochemistry. 
Affinity Purified anti-Huntingtin pS421 antibody was used at a 1:100 dilution to detect phosphorylated Huntingtin by immunohistochemistry in human brain cerebellum. 
Positive cytoplasmic staining is observed in neurons. Tissue was formalin-fixed and paraffin embedded. Personal Communication, Alan Yen, LifeSpanBiosciences, Seattle, WA. Rabbit Aff - Purified Hu E, WB
0.1 mg / €440.00
  Acris Antibodies GmbH

Huntingtin antibody

Total protein extract of normal human cerebral cortex separated as a strip on a 3-12.5% gradient SDS-PAGE gel and Western blotted. The blot was probed with SM1660. Mouse IgG2a HDA3E10 Purified Hu, Ms, Rb C, IP, WB
0.1 mg / €540.00
  Acris Antibodies GmbH

Huntingtin antibody

Total protein extract of normal human cerebral cortex separated as a strip on a 3-
12.5% gradient SDS-PAGE gel and Western blotted. The blot was probed with SM1661. Mouse IgG1 HDB4E10 Purified Hu, Ms, Rb C, IP, WB
0.1 mg / €540.00
  Acris Antibodies GmbH

Huntingtin antibody

Total protein extract of normal human cerebral cortex separated as a strip on a 3- 12.5% gradient SDS-PAGE gel and Western blotted. The blot was probed with SM1662. Mouse IgG1 HDC8A4 Purified Hu, Ms, Rb C, IP, WB
0.1 mg / €540.00
  Acris Antibodies GmbH

Optineurin / HIP7 (115-130) antibody

Adrenal: Formalin-Fixed Paraffin-Embedded (FFPE) Rabbit IgG Aff - Purified Hu, Mky, Prim P, WB
50 µg / €470.00
  Acris Antibodies GmbH

13 Item(s)

per page
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