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GRAF Recombinant Rabbit Monoclonal Antibody [JE62-23]

Usd: 385

Specification

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Catalog# HA722409

GRAF Recombinant Rabbit Monoclonal Antibody [JE62-23]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

GRAF Recombinant Rabbit Monoclonal Antibody [JE62-23]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human GRAF aa 750-814.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Target Molecular Weight

Predicted band size: 92 kDa

Positive Control

Caco-2 cell lysate, Jurkat cell lysate, HepG2 cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, human heart tissue, human brain tissue, mouse brain tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

JE62-23

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

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

  • 1:200-1:1,000

Target

Function

Rho GTPase activating protein 26 (ARHGAP26) also known as GTPase Regulator Associated with Focal Adhesion Kinase (GRAF) is a protein that in humans is encoded by the ARHGAP26 gene. GRAF1 is a multidomain protein that is necessary for the CLIC/GEEC endocytic pathway. By virtue of an N-terminal BAR domain, GRAF1 sculpts the endocytic membranes of this pathway into 40 nm diameter tubules and vesicles that allow uptake of extracellular fluid, GPI-linked proteins and certain bacterial exotoxins into cells. The role of dynamin in the CLIC/GEEC pathway is controversial, but GRAF1 interacts strongly with this protein and acute inhibition of dynamin action abrogates CLIC/GEEC endocytosis. There are several members of the GRAF family of proteins, including GRAF2, GRAF3, and oligophrenin, all of which likely playing similar roles during clathrin-independent endocytic events. Mutations of both GRAF1 and oligophrenin are strongly implicated in causing human disease (leukaemia and mental retardation, respectively). Recently, autoantibodies to ARHGAP26 have been implicated in autoimmune cerebellar ataxia.

Background References

1. Zhang L et al. The role of GTPase-activating protein ARHGAP26 in human cancers. Mol Cell Biochem. 2022 Jan

2. Wang K et al. Associations of ARHGAP26 Polymorphisms with Alzheimer\'s Disease and Cardiovascular Disease. J Mol Neurosci. 2022 May

Subcellular Location

Cell junction, focal adhesion, Cytoplasm, cytoskeleton; Endosome membrane.

UNIPROT

SwissProt: Q9UNA1 Human

SwissProt: Q6ZQ82 Mouse

Entrez Gene: 307459 Rat

Synonyms

arhgap26 antibody

FLJ42530 antibody

GRAF antibody

GRAF1 antibody

GTPase regulator associated with focal adhesion kinase antibody

GTPase regulator associated with focal adhesion kinase pp125(FAK) antibody

KIAA0621 antibody

oligophrenin 1 like protein antibody

Oligophrenin-1-like protein antibody

oligophrenin1like protein antibody

Expand

Images

  • Western blot analysis of GRAF on different lysates with Rabbit anti-GRAF antibody (<a href="/products/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</a>) at 1/1,000 dilution.<br /><br />Lane 1: Caco-2 cell lysate (15 µg/Lane)<br />Lane 2: Jurkat cell lysate (15 µg/Lane)<br />Lane 3: HepG2 cell lysate (15 µg/Lane)<br />Lane 4: MCF7 cell lysate (15 µg/Lane)<br />Lane 5: NIH/3T3 cell lysate (15 µg/Lane)<br />Lane 6: Mouse brain tissue lysate (30 µg/Lane)<br />Lane 7: Rat brain tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 92 kDa<br />Observed band size: 92 kDa<br /><br />Exposure time: 3 minutes; 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/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</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 GRAF on different lysates with Rabbit anti-GRAF antibody (HA722409) at 1/1,000 dilution.

    Lane 1: Caco-2 cell lysate (15 µg/Lane)
    Lane 2: Jurkat cell lysate (15 µg/Lane)
    Lane 3: HepG2 cell lysate (15 µg/Lane)
    Lane 4: MCF7 cell lysate (15 µg/Lane)
    Lane 5: NIH/3T3 cell lysate (15 µg/Lane)
    Lane 6: Mouse brain tissue lysate (30 µg/Lane)
    Lane 7: Rat brain tissue lysate (30 µg/Lane)

    Predicted band size: 92 kDa
    Observed band size: 92 kDa

    Exposure time: 3 minutes; 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 (HA722409) 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-GRAF antibody (<a href="/products/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</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/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</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-GRAF antibody (HA722409) 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 (HA722409) 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 brain tissue with Rabbit anti-GRAF antibody (<a href="/products/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</a>) at 1/1,000 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/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</a>) at 1/1,000 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 brain tissue with Rabbit anti-GRAF antibody (HA722409) at 1/1,000 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 (HA722409) at 1/1,000 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 mouse brain tissue with Rabbit anti-GRAF antibody (<a href="/products/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</a>) at 1/1,000 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/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</a>) at 1/1,000 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 mouse brain tissue with Rabbit anti-GRAF antibody (HA722409) at 1/1,000 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 (HA722409) at 1/1,000 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 rat brain tissue with Rabbit anti-GRAF antibody (<a href="/products/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</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/HA722409" style="font-weight: bold;text-decoration: underline;">HA722409</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 brain tissue with Rabbit anti-GRAF antibody (HA722409) 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 (HA722409) 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.

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