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GEF H1 Recombinant Rabbit Monoclonal Antibody [JG36-46]

Usd: 385

Specification

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Catalog# ET7108-34

GEF H1 Recombinant Rabbit Monoclonal Antibody [JG36-46]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

GEF H1 Recombinant Rabbit Monoclonal Antibody [JG36-46]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human GEF H1 aa 937-986 / 986.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

112 kDa

Positive Control

HeLa cell lysate, NIH/3T3 cell lysate, C6 cell lysate, SH-SY5Y, HUVEC, human tonsil, mouse testis, rat brain tissue.

Conjugation

unconjugated

Clone Number

JG36-46

RRID

AB_3070833

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

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

Target

Function

Activates Rho-GTPases by promoting the exchange of GDP for GTP. May be involved in epithelial barrier permeability, cell motility and polarization, dendritic spine morphology, antigen presentation, leukemic cell differentiation, cell cycle regulation, innate immune response, and cancer. May stimulate instead the cortical activity of Rac. Inactive toward CDC42, TC10, or Ras-GTPases. Forms an intracellular sensing system along with NOD1 for the detection of microbial effectors during cell invasion by pathogens. Required for RHOA and RIP2 dependent NF-kappaB signaling pathways activation upon S.flexneri cell invasion. Involved not only in sensing peptidoglycan (PGN)-derived muropeptides through NOD1 that is independent of its GEF activity, but also in the activation of NF-kappaB by Shigella effector proteins (IpgB2 and OspB) which requires its GEF activity and the activation of RhoA.

Background References

1. Ren Y et al. Cloning and characterization of GEF-H1, a microtubule-associated guanine nucleotide exchange factor for Rac and Rho GTPases. J Biol Chem 273:34954-34960 (1998).

2. Fukazawa A et al. GEF-H1 mediated control of NOD1 dependent NF-kappaB activation by Shigella effectors. PLoS Pathog 4:E1000228-E1000228 (2008).

Post-translational Modification

Phosphorylation of Ser-886 by PAK1 induces binding to protein YWHAZ, promoting its relocation to microtubules and the inhibition of its activity. Phosphorylated by AURKA and CDK1 during mitosis, which negatively regulates its activity. Phosphorylation by MAPK1 or MAPK3 increases nucleotide exchange activity. Phosphorylation by PAK4 releases GEF-H1 from the microtubules. Phosphorylated on serine, threonine and tyrosine residues in a RIPK2-dependent manner.

Subcellular Location

Ruffle membrane, cytoskeleton, spindle, Golgi apparatus, Cytoplasm, tight junction, Cytoplasmic vesicle.

UNIPROT

SwissProt: Q92974 Human

SwissProt: Q60875 Mouse

SwissProt: Q5FVC2 Rat

Synonyms

AA408978 antibody

ARHG2 antibody

ARHG2_HUMAN antibody

ARHGEF 2 antibody

ARHGEF-2 antibody

ARHGEF2 antibody

GEF antibody

GEF H1 antibody

GEF-H1 antibody

GEFH1 antibody

Expand

Images

  • Western blot analysis of GEF H1 on different lysates with Rabbit anti-GEF H1 antibody (<a href="/products/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: NIH/3T3 cell lysate<br />Lane 3: C6 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br />Exposure time: 3 minutes; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 112 kDa<br />Observed band size: 112 kDa

    Western blot analysis of GEF H1 on different lysates with Rabbit anti-GEF H1 antibody (ET7108-34) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: NIH/3T3 cell lysate
    Lane 3: C6 cell lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 3 minutes; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET7108-34, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 112 kDa
    Observed band size: 112 kDa

  • Immunocytochemistry analysis of SH-SY5Y cells labeling GEF H1 with Rabbit anti-GEF H1 antibody (<a href="/products/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-GEF H1 antibody (<a href="/products/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of SH-SY5Y cells labeling GEF H1 with Rabbit anti-GEF H1 antibody (ET7108-34) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-GEF H1 antibody (ET7108-34) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

  • Immunocytochemistry analysis of HUVEC cells labeling GEF H1 with Rabbit anti-GEF H1 antibody (<a href="/products/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-GEF H1 antibody (<a href="/products/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HUVEC cells labeling GEF H1 with Rabbit anti-GEF H1 antibody (ET7108-34) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-GEF H1 antibody (ET7108-34) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded human tonsil with Rabbit anti-GEF H1 antibody (<a href="/products/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution.<br /><br />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 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/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution for 0.5 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 tonsil with Rabbit anti-GEF H1 antibody (ET7108-34) at 1/50 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-34) at 1/50 dilution for 0.5 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 testis with Rabbit anti-GEF H1 antibody (<a href="/products/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution.<br /><br />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 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/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution for 0.5 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 testis with Rabbit anti-GEF H1 antibody (ET7108-34) at 1/50 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-34) at 1/50 dilution for 0.5 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-GEF H1 antibody (<a href="/products/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution.<br /><br />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 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/ET7108-34" style="font-weight: bold;text-decoration: underline;">ET7108-34</a>) at 1/50 dilution for 0.5 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-GEF H1 antibody (ET7108-34) at 1/50 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-34) at 1/50 dilution for 0.5 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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