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STUB1 Recombinant Rabbit Monoclonal Antibody [JG38-22]

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Specification

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Catalog# ET7108-65

STUB1 Recombinant Rabbit Monoclonal Antibody [JG38-22]

Application

  • WB

  • IF-Cell

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

STUB1 Recombinant Rabbit Monoclonal Antibody [JG38-22]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human STUB1 aa 10-59 / 303.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, IP, FC

Molecular Weight

Predicted band size: 35 kDa

Positive Control

HeLa cell lysate, HEK-293 cell lysate, human brain tissue lysate, human heart tissue, mouse heart tissue, rat heart tissue, 293T, LOVO, MCF-7, SH-SY5Y.

Conjugation

unconjugated

Clone Number

JG38-22

RRID

AB_3070861

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

Target

Function

CHIP (carboxy terminus of HSP 70-interacting protein), also designated STIP1 homology and U-box containing protein 1, HSPABP2, NY-CO-7, SDCCAG7 and STUB1, is a cytoplasmic E3 ubiquitin ligase that influences protein ubiquitylation. CHIP interacts with Smad1/Smad4 and blocks BMP signaling through the ubiquitin-mediated degradation of Smad proteins. CHIP controls both association of Hsp70/Hsp90 chaperones with ErbB2 and down-regulation of ErbB2 induced by inhibitors of Hsp90. A 1.3-kb transcript is most abundant in striated muscle (heart and skeletal muscle), with lower expression in pancreas and brain.

Background References

1. Ballinger C A et al. Identification of CHIP, a novel tetratricopeptide repeat-containing protein that interacts with heat shock proteins and negatively regulates chaperone functions. Mol Cell Biol 19:4535-4545 (1999).

2. Jiang J et al. CHIP is a U-box-dependent E3 ubiquitin ligase: identification of Hsc70 as a target for ubiquitylation. J Biol Chem 276:42938-42944 (2001).

Tissue Specificity

Expressed in differentiated myotubes (at protein level). Highly expressed in skeletal muscle, heart, pancreas, brain and placenta. Detected in kidney, liver and lung.

Post-translational Modification

Monoubiquitinated at Lys-2 following cell stress by UBE2W, promoting the interaction with ATXN3 (By similarity). Auto-ubiquitinated; mediated by UBE2D1 and UBE2D2.

Subcellular Location

Nucleus. Cytoplasm.

UNIPROT

SwissProt: Q9UNE7 Human

SwissProt: Q9WUD1 Mouse

Entrez Gene: 287155 Rat

Synonyms

Antigen NY CO 7 antibody

Antigen NY-CO-7 antibody

C terminus of Hsp70-interacting protein antibody

Carboxy terminus of Hsp70 interacting protein antibody

Carboxy terminus of Hsp70-interacting protein antibody

Carboxy terminus of Hsp70p interacting protein antibody

CHIP antibody

CHIP_HUMAN antibody

CLL associated antigen KW 8 antibody

CLL-associated antigen KW-8 antibody

Expand

Images

  • Western blot analysis of STUB1 on different lysates with Rabbit anti-STUB1 antibody (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (15 µg/Lane)<br />Lane 2: HEK-293 cell lysate (15 µg/Lane)<br />Lane 3: Human brain tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 35 kDa<br />Observed band size: 34 kDa<br /><br />Exposure time: 14 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of STUB1 on different lysates with Rabbit anti-STUB1 antibody (ET7108-65) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (15 µg/Lane)
    Lane 2: HEK-293 cell lysate (15 µg/Lane)
    Lane 3: Human brain tissue lysate (30 µg/Lane)

    Predicted band size: 35 kDa
    Observed band size: 34 kDa

    Exposure time: 14 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 (ET7108-65) at 1/1,000 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.

  • Immunohistochemical analysis of paraffin-embedded human heart tissue with Rabbit anti-STUB1 antibody (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>) at 1/200 dilution.<br /><br />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 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>) at 1/200 dilution for 1 hour 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.

    Immunohistochemical analysis of paraffin-embedded human heart tissue with Rabbit anti-STUB1 antibody (ET7108-65) at 1/200 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-65) at 1/200 dilution for 1 hour 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.

  • Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-STUB1 antibody (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>) at 1/200 dilution.<br /><br />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 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>) at 1/200 dilution for 1 hour 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.

    Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-STUB1 antibody (ET7108-65) at 1/200 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-65) at 1/200 dilution for 1 hour 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.

  • Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-STUB1 antibody (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>) at 1/200 dilution.<br /><br />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 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>) at 1/200 dilution for 1 hour 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.

    Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-STUB1 antibody (ET7108-65) at 1/200 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-65) at 1/200 dilution for 1 hour 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.

  • ICC staining of STUB1 in 293T 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 (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of STUB1 in 293T 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 (ET7108-65, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of STUB1 in LOVO 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 (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of STUB1 in LOVO 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 (ET7108-65, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of STUB1 in MCF-7 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 (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of STUB1 in MCF-7 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 (ET7108-65, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Flow cytometric analysis of STUB1 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET7108-65" style="font-weight: bold;text-decoration: underline;">ET7108-65</a>, 1/50) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; yellow).

    Flow cytometric analysis of STUB1 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7108-65, 1/50) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; yellow).

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Citation