PRODUCT CODE: EM1901-04

Hip1 Monoclonal Antibody (EM1901-04)

Applications

  • WB

  • ICC

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Hip1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: SH-SY5Y cell lysate<br />
 Lane 2: A549 cell lysate
  • Western blot analysis of Hip1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: SH-SY5Y cell lysate<br />
 Lane 2: A549 cell lysate
  • ICC staining of Hip1 in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1901-04, 1/50 dilution) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Hip1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-04, 1/200 dilution) for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-Hip1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with primary antibody (EM1901-04, 1/200 dilution) for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue using anti-Hip1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with primary antibody (EM1901-04, 1/200 dilution) for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Hip1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with primary antibody (EM1901-04, 1/200 dilution) for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
Western blot analysis of Hip1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: SH-SY5Y cell lysate
Lane 2: A549 cell lysate

Applications

  • WB

  • ICC

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

Hip1 Monoclonal Antibody (EM1901-04)

Immunogen

Synthetic peptide within human hip1 aa 8-31.

Host

Mouse

Positive Control

SH-SY5Y, A549, rat brain tissue, human colon cancer tissue, human prostate cancer tissue, mouse brain tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

13E1

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Peptide affinity purified.

MOLECULAR WEIGHT

116 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB:1:500-1:2,000

  • ICC:1:50-1:200

  • IHC-P:1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Hip1

SYNONYMS

2610109B09Rik antibody; A930014B11Rik antibody; E130315I21Rik antibody; HIP 1 antibody; HIP I antibody; HIP-1 antibody; HIP-I antibody; hip1 antibody; HIP1/PDGFRB fusion gene antibody; HIP1/PDGFRB fusion gene, included antibody; HIP1_HUMAN antibody; HIPI antibody; Huntingtin interacting protein 1 antibody; Huntingtin Interacting Protein HIP1 antibody; Huntingtin-interacting protein 1 antibody; Huntingtin-interacting protein I antibody; ILWEQ antibody; KIAA4113 antibody; MGC126506 antibody; MGC27616 antibody; mKIAA4113 antibody

SEQUENCE SIMILARITIES

Belongs to the SLA2 family.

TISSUE SPECIFICITY

Ubiquitously expressed with the highest level in brain. Expression is up-regulated in prostate and colon cancer.

SUBCELLULAR LOCATION

Nucleus. Cytoplasm.

FUNCTION

Huntington disease is associated with the expansion of a polyglutamine tract, greater than 35 repeats, in the HD gene product huntingtin. HIP1 (huntingtin-interacting protein 1), a membrane-associated protein, binds specifically to the N-terminus of human huntingtin. HIP1 is ubiquitously expressed in different brain regions at low levels, and exhibits nearly identical subcellular fractionation as huntingtin. The huntingtin-HIP1 interaction is restricted to the brain and is inversely correlated to the polyglutamine length in the huntingtin, suggesting that loss of normal huntingtin-HIP1 interaction may compromise the membrane-cytoskeletal integrity in the brain. HIP1 contains an endocytic multidomain protein with a C-terminal Actin-binding domain, a central coiled-coil forming region and an N-terminal ENTH domain. HIP1 may be involved in vesicle trafficking; the structural integrity of HIP1 is crucial for maintenance of normal vesicle size in vivo. HIP12 is a non-proapoptotic member of the HIP gene family that is expressed in the brain and shares a similar subcellular distribution pattern with HIP1. However, HIP12 differs from HIP1 in its pattern of expression at both the mRNA and protein level. HIP12 does not directly interact with huntingtin but can interact with HIP1.