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Rac1-2-3 Recombinant Rabbit Monoclonal Antibody [JM11-29]

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Specification

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Catalog# ET1703-80

Rac1-2-3 Recombinant Rabbit Monoclonal Antibody [JM11-29]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Rac1-2-3 Recombinant Rabbit Monoclonal Antibody [JM11-29]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant full length protein of Human RAC1 aa 1-192 / 192.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P

Target Molecular Weight

Predicted band size: 21 kDa

Positive Control

MCF-7 cell lysate, PC-12 cell lysate, 293T, Hela, NIH/3T3, human spleen tissue, mouse brain tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

JM11-29

RRID

AB_3070435

Reactivity Data

WB IF-Cell IF-Tissue IHC-P
Human
Tested Tested
Tested Tested
Tested Tested
Mouse
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Rat
Tested Tested
Tested Tested

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

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:200

  • IHC-P

  • 1:200

Target

Function

Plasma membrane-associated small GTPase which cycles between active GTP-bound and inactive GDP-bound states. In its active state, binds to a variety of effector proteins to regulate cellular responses such as secretory processes, phagocytosis of apoptotic cells, epithelial cell polarization and growth-factor induced formation of membrane ruffles. Rac1 p21/rho GDI heterodimer is the active component of the cytosolic factor sigma 1, which is involved in stimulation of the NADPH oxidase activity in macrophages (By similarity). Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. Isoform B has an accelerated GEF-independent GDP/GTP exchange and an impaired GTP hydrolysis, which is restored partially by GTPase-activating proteins. It is able to bind to the GTPase-binding domain of PAK but not full-length PAK in a GTP-dependent manner, suggesting that the insertion does not completely abolish effector interaction.

Background References

1. Yoshida S et al. Growth factor signaling to mTORC1 by amino acid-laden macropinosomes. J Cell Biol 211:159-72 (2015).

2. Skvortsov S et al. Rac1 as a potential therapeutic target for chemo-radioresistant head and neck squamous cell carcinomas (HNSCC). Br J Cancer 110:2677-87 (2014).

Sequence Similarity

Belongs to the small GTPase superfamily. Rho family.

Tissue Specificity

Isoform B is predominantly identified in skin and epithelial tissues from the intestinal tract. Its expression is elevated in colorectal tumors at various stages of neoplastic progression, as compared to their respective adjacent tissues.

Post-translational Modification

(Microbial infection) AMPylation at Tyr-32 and Thr-35 are mediated by bacterial enzymes in case of infection by H.somnus and V.parahaemolyticus, respectively. AMPylation occurs in the effector region and leads to inactivation of the GTPase activity by preventing the interaction with downstream effectors, thereby inhibiting actin assembly in infected cells. It is unclear whether some human enzyme mediates AMPylation; FICD has such ability in vitro but additional experiments remain to be done to confirm results in vivo.; GTP-bound active form is ubiquitinated by HACE1, leading to its degradation by the proteasome.; (Microbial infection) Glycosylated at Tyr-32 by Photorhabdus asymbiotica toxin PAU_02230. Mono-O-GlcNAcylation by PAU_02230 inhibits downstream signaling by an impaired interaction with diverse regulator and effector proteins of Rac and leads to actin disassembly.

Subcellular Location

Cell membrane, Cell projection, Cytoplasm, Cytoskeleton, Membrane.

UNIPROT

SwissProt: P15153 Human

SwissProt: P60763 Human

SwissProt: P63000 Human

SwissProt: P60764 Mouse

SwissProt: P63001 Mouse

SwissProt: Q05144 Mouse

SwissProt: Q6RUV5 Rat

SwissProt: Q5U1Y2 Rat

SwissProt: M0R5T4 Rat

Synonyms

Cell migration inducing gene 5 protein antibody

EN 7 antibody

OTTMUSP00000004488 antibody

p21 Rac3 antibody

p21-Rac3 antibody

Rac1B antibody

RAC3 antibody

RAC3_HUMAN antibody

RAS related C3 botulinum substrate 3 antibody

Ras related C3 botulinum toxin substrate 3 (rho family small GTP binding protein Rac3) antibody

Expand

Images

  • Western blot analysis of Rac1-2-3 on MCF-7 cell lysates with Rabbit anti-Rac1-2-3 antibody (<a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</a>) at 1/2,000 dilution.<br /><br /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 1 minutes; ECL: K1801<br /><br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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: 21.5kDa<br />Observed band size: 21kDa

    Western blot analysis of Rac1-2-3 on MCF-7 cell lysates with Rabbit anti-Rac1-2-3 antibody (ET1703-80) at 1/2,000 dilution.


    Lysates/proteins at 10 µg/Lane.
    Exposure time: 1 minutes; ECL: K1801


    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1703-80, 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: 21.5kDa
    Observed band size: 21kDa

  • Western blot analysis of Rac1-2-3 on mouse brain tissue lysates with Rabbit anti-Rac1-2-3 antibody (<a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</a>) at 1/2,000 dilution.<br /><br /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 1 minutes; ECL: K1801<br /><br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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: 21.5kDa<br />Observed band size: 21kDa

    Western blot analysis of Rac1-2-3 on mouse brain tissue lysates with Rabbit anti-Rac1-2-3 antibody (ET1703-80) at 1/2,000 dilution.


    Lysates/proteins at 10 µg/Lane.
    Exposure time: 1 minutes; ECL: K1801


    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1703-80, 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: 21.5kDa
    Observed band size: 21kDa

  • ICC staining of Rac1-2-3 in 293T cells (red). 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/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;594 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 Rac1-2-3 in 293T cells (red). 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 (ET1703-80, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 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 Rac1-2-3 in Hela cells (red). 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/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;594 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 Rac1-2-3 in Hela cells (red). 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 (ET1703-80, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 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 Rac1-2-3 in NIH/3T3 cells (red). 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/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;594 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 Rac1-2-3 in NIH/3T3 cells (red). 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 (ET1703-80, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 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 human spleen tissue with Rabbit anti-Rac1-2-3 antibody (<a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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 spleen tissue with Rabbit anti-Rac1-2-3 antibody (ET1703-80) 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 (ET1703-80) 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 brain tissue with Rabbit anti-Rac1-2-3 antibody (<a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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 brain tissue with Rabbit anti-Rac1-2-3 antibody (ET1703-80) 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 (ET1703-80) 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-Rac1-2-3 antibody (<a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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-Rac1-2-3 antibody (ET1703-80) 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 (ET1703-80) 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.

  • Immunofluorescence analysis of paraffin-embedded mouse embryonic brain tissue labeling Rac1-2-3 with Rabbit anti-Rac1-2-3 antibody (<a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</a>, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. 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. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of paraffin-embedded mouse embryonic brain tissue labeling Rac1-2-3 with Rabbit anti-Rac1-2-3 antibody (ET1703-80) 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1703-80, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  • Immunofluorescence analysis of paraffin-embedded mouse embryonic eye tissue labeling Rac1-2-3 with Rabbit anti-Rac1-2-3 antibody (<a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/ET1703-80" style="font-weight: bold;text-decoration: underline;">ET1703-80</a>, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. 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. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of paraffin-embedded mouse embryonic eye tissue labeling Rac1-2-3 with Rabbit anti-Rac1-2-3 antibody (ET1703-80) 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1703-80, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Citation

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