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GRB7 Recombinant Rabbit Monoclonal Antibody [JE50-05]

Usd: 385

Specification

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Catalog# HA721924

GRB7 Recombinant Rabbit Monoclonal Antibody [JE50-05]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

GRB7 Recombinant Rabbit Monoclonal Antibody [JE50-05]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human GRB7 aa 1-150 / 532.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 60 kDa

Positive Control

A431 cell lysate, HepG2 cell lysate, SK-Br-3 cell lysate, human breast cancer tissue, human kidney tissue, SK-Br-3.

Conjugation

unconjugated

Clone Number

JE50-05

RRID

AB_3096583

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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

  • IHC-P

  • 1:500-1:2,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

Adapter protein that interacts with the cytoplasmic domain of numerous receptor kinases and modulates down-stream signaling. Promotes activation of down-stream protein kinases, including STAT3, AKT1, MAPK1 and/or MAPK3. Promotes activation of HRAS. Plays a role in signal transduction in response to EGF. Plays a role in the regulation of cell proliferation and cell migration. Plays a role in the assembly and stability of RNA stress granules. Binds to the 5'UTR of target mRNA molecules and represses translation of target mRNA species, when not phosphorylated. Phosphorylation impairs RNA binding and promotes stress granule disassembly during recovery after cellular stress.

Background References

1. Han D.C., Shen T.L., Guan J.L. Role of Grb7 targeting to focal contacts and its phosphorylation by focal adhesion kinase in regulation of cell migration. J. Biol. Chem. 275:28911-28917 (2000).

2. Shen T.L., Han D.C., Guan J.L. Association of Grb7 with phosphoinositides and its role in the regulation of cell migration. J. Biol. Chem. 277:29069-29077 (2002).

Subcellular Location

Cell junction. Cell membrane.

UNIPROT

SwissProt: Q14451 Human

Synonyms

B47 antibody

Epidermal growth factor receptor GRB 7 antibody

Epidermal growth factor receptor GRB-7 antibody

GRB7 adapter protein antibody

GRB7 antibody

GRB7_HUMAN antibody

Growth factor receptor bound protein 7 antibody

Growth factor receptor-bound protein 7 antibody

OTTHUMP00000164349 antibody

OTTHUMP00000164350 antibody

Expand

Images

  • Western blot analysis of GRB7 on different lysates with Rabbit anti-GRB7 antibody (<a href="/products/HA721924" style="font-weight: bold;text-decoration: underline;">HA721924</a>) at 1/1,000 dilution.<br /><br />Lane 1: A431 cell lysate (40 µg/Lane)<br />Lane 2: HepG2 cell lysate (40 µg/Lane)<br />Lane 3: SK-Br-3 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 60 kDa<br />Observed band size: 60 kDa<br /><br />Exposure time: Lane 1-2: 1 minute 40 seconds; Lane 3: 3 minutes;<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/HA721924" style="font-weight: bold;text-decoration: underline;">HA721924</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 GRB7 on different lysates with Rabbit anti-GRB7 antibody (HA721924) at 1/1,000 dilution.

    Lane 1: A431 cell lysate (40 µg/Lane)
    Lane 2: HepG2 cell lysate (40 µg/Lane)
    Lane 3: SK-Br-3 cell lysate (20 µg/Lane)

    Predicted band size: 60 kDa
    Observed band size: 60 kDa

    Exposure time: Lane 1-2: 1 minute 40 seconds; Lane 3: 3 minutes;

    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 (HA721924) 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 breast cancer tissue with Rabbit anti-GRB7 antibody (<a href="/products/HA721924" style="font-weight: bold;text-decoration: underline;">HA721924</a>) at 1/2,000 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/HA721924" style="font-weight: bold;text-decoration: underline;">HA721924</a>) at 1/2,000 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 breast cancer tissue with Rabbit anti-GRB7 antibody (HA721924) at 1/2,000 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 (HA721924) at 1/2,000 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 kidney tissue with Rabbit anti-GRB7 antibody (<a href="/products/HA721924" style="font-weight: bold;text-decoration: underline;">HA721924</a>) at 1/2,000 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/HA721924" style="font-weight: bold;text-decoration: underline;">HA721924</a>) at 1/2,000 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 kidney tissue with Rabbit anti-GRB7 antibody (HA721924) at 1/2,000 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 (HA721924) at 1/2,000 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.

  • Immunocytochemistry analysis of SK-Br-3 cells labeling GRB7 with Rabbit anti-GRB7 antibody (<a href="/products/HA721924" style="font-weight: bold;text-decoration: underline;">HA721924</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GRB7 antibody (<a href="/products/HA721924" style="font-weight: bold;text-decoration: underline;">HA721924</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of SK-Br-3 cells labeling GRB7 with Rabbit anti-GRB7 antibody (HA721924) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GRB7 antibody (HA721924) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

  • Flow cytometric analysis of SK-Br-3 cells labeling GRB7.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721924" style="font-weight: bold;text-decoration: underline;">HA721924</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of SK-Br-3 cells labeling GRB7.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721924, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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