PRODUCT CODE: ET7108-65

STUB1 Recombinant Rabbit Monoclonal Antibody [JG38-22] (ET7108-65)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of STUB1 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 (ET7108-65, 1/500) was used 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: 293 cell lysate<br />
Lane 2: MCF-7 cell lysate<br />
Lane 3: SK-Br-3 cell lysate
  • Western blot analysis of STUB1 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 (ET7108-65, 1/500) was used 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: 293 cell lysate<br />
Lane 2: MCF-7 cell lysate<br />
Lane 3: SK-Br-3 cell lysate
  • 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 (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 (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).
  • Immunohistochemical analysis of paraffin-embedded rat epididymis tissue using anti-STUB1 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 (ET7108-65, 1/50) for 30 minutes 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 colon carcinoma tissue using anti-STUB1 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 (ET7108-65, 1/50) for 30 minutes 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 placenta tissue using anti-STUB1 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 (ET7108-65, 1/50) for 30 minutes 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 kidney tissue using anti-STUB1 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 (ET7108-65, 1/50) for 30 minutes 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.
  • 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).
Western blot analysis of STUB1 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 (ET7108-65, 1/500) was used 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: 293 cell lysate
Lane 2: MCF-7 cell lysate
Lane 3: SK-Br-3 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

STUB1 Recombinant Rabbit Monoclonal Antibody [JG38-22] (ET7108-65)

Immunogen

Recombinant protein within human stub1 aa 1-200.

Host

Rabbit

Positive Control

293 cell lysate, MCF-7 cell lysate, SK-Br-3 cell lysate, 293T, LOVO, MCF-7, rat epididymis tissue, human colon carcinoma tissue, human placenta tissue, mouse kidney tissue, SH-SY5Y.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JG38-22

PROPERTIES

Form

Liquid

Storage Condition

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

Storage Buffer

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

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

35 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • 1:10-1:50

TARGET

UNIPROT #

PROTEIN NAME

STUB1

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; E3 ubiquitin protein ligase CHIP antibody; E3 ubiquitin-protein ligase CHIP antibody; Heat shock protein A binding protein 2 (c terminal) antibody; HSPABP2 antibody; NY CO 7 antibody; PP1131 antibody; SDCCAG7 antibody; Serologically defined colon cancer antigen 7 antibody; STIP1 homology and U Box containing protein 1 antibody; STIP1 homology and U box containing protein 1 E3 ubiquitin protein ligase antibody; STIP1 homology and U box-containing protein 1 antibody; STUB 1 antibody; STUB1 antibody; UBOX 1 antibody; UBOX1 antibody

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.

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.