Overview

  • Product nameAnti-Rac1 antibody [0.T.127]See all Rac1 primary antibodies ...
  • Description
    Mouse monoclonal [0.T.127] to Rac1
  • SpecificityRecognizes Rac at 21kD.Slight Rac 2 cross-reactivity
  • Tested applicationsIHC-Fr, WB, Flow Cyt, IHC (PFA fixed), ICC/IF, ELISA, IP, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Human
  • Immunogen

    Recombinant protein containing the full length human Rac.

  • Positive control
    • WB/IP: rat brain microsomal protein preparation IHC: rat brain sections fixed first with PBS, 4% paraformaldehyde, 2% acetic acid for 30 minutes followed by ethanol:acetic acid [95:5] for 2 minutes. Affinity Precipitation Assay: lysates of 3T3 fibroblasts

Properties

Applications

Our Abpromise guarantee covers the use of ab33186 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 21 kDa.
Flow Cyt Use at an assay dependent concentration.
IHC (PFA fixed) 1/10 - 1/50.
ICC/IF 1/10 - 1/50.
ELISA 1/100 - 1/1000.
IP Use 1µg for 107 cells.
IHC-P Use at an assay dependent concentration.

Target

  • FunctionPlasma 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.
  • Tissue specificityIsoform 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.
  • Sequence similaritiesBelongs to the small GTPase superfamily. Rho family.
  • DomainThe effector region mediates interaction with DEF6.
  • Post-translational
    modifications
    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.
  • Cellular localizationCell membrane. Melanosome. Inner surface of plasma membrane possibly with attachment requiring prenylation of the C-terminal cysteine (By similarity). Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Target information above from: UniProt accession P63000 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • Cell migration inducing gene 5 protein antibody
    • Cell migration-inducing gene 5 protein antibody
    • MGC111543 antibody
    • MIG 5 antibody
    • MIG5 antibody
    • Migration inducing gene 5 antibody
    • Migration inducing protein 5 antibody
    • p21 Rac1 antibody
    • p21-Rac1 antibody
    • Rac 1 antibody
    • RAC1 antibody
    • RAC1_HUMAN antibody
    • Ras like protein TC25 antibody
    • Ras related C3 botulinum toxin substrate 1 (rho family, small GTP binding protein Rac1) antibody
    • Ras-like protein TC25 antibody
    • Ras-related C3 botulinum toxin substrate 1 antibody
    • rho family small GTP binding protein Rac1 antibody
    • TC 25 antibody
    • TC25 antibody
    see all

Anti-Rac1 antibody [0.T.127] images

  • Anti-Rac1 antibody [0.T.127] (ab33186) at 1/750 dilution (for 14 hours at 4°C) + Mouse brain whole tissue lysate at 15 µg

    Secondary
    An HRP-conjugated Goat anti-mouse IgG polyclonal at 1/10000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa
    Observed band size : 21 kDa


    Exposure time : 2 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ab33186 staining Rac1 in Mouse intestine tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% BSA 1 hour at 20°C ; antigen retrieval was by heat mediation with citric acid. Samples were incubated with primary antibody (1/750) for 4 hours at 20°C. An HRP-conjugated Goat anti-mouse IgG polyclonal (1/5000) was used as the secondary antibody.

    See Abreview

References for Anti-Rac1 antibody [0.T.127] (ab33186)

This product has been referenced in:

See all 9 Publications for this product

Product Wall

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (12% acrylamide gel)
Sample Human Tissue lysate - whole (Brain)
Specification Brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Sep 24 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Milk as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step Enzymatic
Sample Rat Tissue sections (GONAD)
Specification GONAD
Permeabilization No
Fixative Formaldehyde
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Submitted Sep 23 2013

Thank you for contacting us.


This antibody has not been epitope mapped.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Vielen Dank für Ihre Anfrage. Gegen die genannten NADPH-Oxidase-Komponenten haben wir folgende Antikörper bzw. Proteine in unserem Katalog: Click here (or use the following: http://www.abcam.com/index.html?datasheet=109366). Click here (or use the...

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Thank you for taking time to complete the questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring...

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Thank you for contacting us. The other positive control lysates you can use are Hela, MCF7, A431 or Raji cell lysates, or Human tubular colorectal, prostate and squamous cells carcinoma tissue lysates. I hope this information is helpful ...

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Application Western blot
Sample Human Cell lysate - whole cell (HCT116)
Loading amount 50 µg
Specification HCT116
Gel Running Conditions Reduced Denaturing (gradient 4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 18°C
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Submitted Jun 29 2011

Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton x-100 10min
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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Submitted Dec 09 2010

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Mouse Intestine)
Specification Mouse Intestine
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Sep 01 2010

Application Western blot
Sample Mouse Tissue lysate - whole (Mouse Brain)
Loading amount 15 µg
Specification Mouse Brain
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Aug 17 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"