Overview

  • Product name
    Anti-Rac1 antibody [23A8]
    See all Rac1 primary antibodies
  • Description
    Mouse monoclonal [23A8] to Rac1
  • Host species
    Mouse
  • Specificity
    Recognizes Rac at 21kD.Slight Rac 2 cross-reactivity. The clone number has been updated from (0.T.127) to (23A8) both clone numbers name the same antibody clone.
  • Tested applications
    Suitable for: IHC-Fr, WB, Flow Cyt, IHC (PFA fixed), ICC/IF, ELISA, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Human
  • Immunogen

    Recombinant full length protein corresponding to Human Rac1.

  • Positive control
    • WB/IP: rat brain microsomal protein preparation, Mouse brain IHC-P: Mouse intestine tissue
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. 

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 Abreviews Notes
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 0.3 - 2 µg/ml. Predicted molecular weight: 21 kDa.
Flow Cyt Use at an assay dependent concentration.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

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

  • 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.
  • 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.
  • Sequence similarities
    Belongs to the small GTPase superfamily. Rho family.
  • Domain
    The 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 localization
    Cell 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.
  • Information by UniProt
  • Database links
  • Form
    Rac1 has high sequence homology with other Rac proteins. Therefore some antibodies may show cross-reactivity.
  • Alternative names
    • Cell migration inducing gene 5 protein antibody
    • Cell migration-inducing gene 5 protein antibody
    • MGC111543 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

Images

  • Anti-Rac1 antibody [23A8] (ab33186) at 0.3 µg/ml + Rat Brain Microsomes at 40 µg

    Predicted band size: 21 kDa

  • 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

  • Anti-Rac1 antibody [23A8] (ab33186) at 1/750 dilution (for 14 hours at 4°C) + Mouse brain 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


    Blocking Step: 5% Milk for 1 hour at 22°C

    See Abreview

References

This product has been referenced in:
  • Liang X  et al. Reactive oxygen species mediated oxidative stress links diabetes and atrial fibrillation. Mol Med Rep 17:4933-4940 (2018). Read more (PubMed: 29393403) »
  • Lalowski MM  et al. Characterizing the Key Metabolic Pathways of the Neonatal Mouse Heart Using a Quantitative Combinatorial Omics Approach. Front Physiol 9:365 (2018). Read more (PubMed: 29695975) »
See all 43 Publications for this product

Customer reviews and Q&As

1-10 of 26 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (fibroblast)
Permeabilization
Yes - 0.3% Triton X-100 in blocking buffer
Specification
fibroblast
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 30 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
30 µg
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 21 2017

Application
Western blot
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 01 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 01 2016

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (16%)
Sample
Mouse Cell lysate - whole cell (Primary bone marrow derived macrophage)
Specification
Primary bone marrow derived macrophage
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 16 2014

Answer



With regards to the specificity of this antibody, its immunogen is recombinant protein containing the full length human Rac. Unfortunately we have not determined the epitope and therefore do not know to exactly whihc part of the protein it binds to. Rac1b difers to rac1a in that it has extra amino acids inserted at amino acid position 75: T → TVGETYGKDITSRGKDKPIA. If the antibody does not bind on or around amino acid 75, then it is very possible that it also recognises isoform rac1b which has a molecular weight of 24 kDa and could explain a band at a higher molecular weight. At what molecular weight do you observe this higher band at?

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Sep 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Sep 23 2013

Answer

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.

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Answer

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