Anti-RAC1 + Cdc42 (phospho S71) antibody (ab5482)

Overview

  • Product name
    Anti-RAC1 + Cdc42 (phospho S71) antibody
    See all RAC1 + Cdc42 primary antibodies
  • Description
    Rabbit polyclonal to RAC1 + Cdc42 (phospho S71)
  • Specificity
    Cdc 42 [pS71] (100% homologous) and Rho A/B/C [pS73] (92% homologous) are expected to react.
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic phospho peptide (Human) containing serine 71. The sequence is conserved in human and mouse RAC 1, 2, and 3, and Cdc 42 human, mouse, rat, dog and frog.

  • Positive control
    • A431 cells treated with EGF.
  • General notes


    RAC, Cdc 42 and Rho A, B, and C are members of a small RhoGTPase family that bind and hydrolyze GTP. GTP bound RAC 1 and cdc 42 play a pivotal role in controlling cell shape, adhesion, growth and transformation. Active Rac 1 is implicated in regulating serum response element (SRE), NFAT 1 and nuclear factor kappa B (NF kappa B) transcription activities. Activated RAC 1 and Cdc 42 bind and activate PAK 1, which in turn activates key downstream signaling proteins including MEKK 1 and JNK. RAC 1 and Cdc 42 are phosphorylated on serine 71, a putative Akt site located between the protein binding domain and GTP binding domain. Phosphorylation of RAC 1 on serine 71 regulates its GTP binding and GTPase activity.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.05% Sodium Azide
    Constituents: 50% Glycerol, PBS, 1mg/ml BSA (IgG, protease free). pH 7.3
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated RAC 1. The final product is generated by affinity chromatography using a RAC 1 derived peptide that is phosphorylated at serine 71.
  • Primary antibody notes
    RAC, Cdc 42 and Rho A, B, and C are members of a small RhoGTPase family that bind and hydrolyze GTP. GTP bound RAC 1 and cdc 42 play a pivotal role in controlling cell shape, adhesion, growth and transformation. Active Rac 1 is implicated in regulating serum response element (SRE), NFAT 1 and nuclear factor kappa B (NF kappa B) transcription activities. Activated RAC 1 and Cdc 42 bind and activate PAK 1, which in turn activates key downstream signaling proteins including MEKK 1 and JNK. RAC 1 and Cdc 42 are phosphorylated on serine 71, a putative Akt site located between the protein binding domain and GTP binding domain. Phosphorylation of RAC 1 on serine 71 regulates its GTP binding and GTPase activity.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5482 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
WB 1/1000. Detects a band of approximately 23 kDa.

Target

  • Relevance
    Cdc42/Rac belongs to the superfamily of small GTPases that are structurally linked to the proto-oncogene product p21ras and are important for the control of cell growth and differentiation as well as for intracellular organization. Cdc42/Rac is an important upstream regulator of the protein kinase cascade that controls the SAPK/JNK and p38 activity. Recent data also suggest that constitutive active forms of Cdc42 can induce apoptosis through a mechanism requiring signaling through SAPK/JNK.
  • Cellular localization
    Cell membrane; Lipid anchor; Cytoplasmic side.
  • Database links
  • Alternative names
    • CDC42Hs antibody
    • Cell division cycle 42 antibody
    • G25K antibody
    • MIG5 antibody
    • Migration inducing gene 5 antibody
    • p21 Rac1 antibody
    • Ras like protein TC25 antibody
    • Small GTP binding protein Cdc42 antibody
    • TC25 antibody
    see all

Images

  • Peptide Competition and Phosphatase Treatment:
    Lysates prepared from A431 cells left unstimulated (1) or stimulated with EGF (2-6) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either not treated (1-5) or treated with lambda phosphatase (6), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ab5482 antibody for two hours at room temperature in a 3% BSATBST buffer, following prior incubation with: no peptide (1, 2, 6), the nonphosphopeptide corresponding to the immunogen (3), a generic phosphoserine containing peptide (4), or, the phosphopeptide immunogen (5), After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that the peptide corresponding to ab5482 blocks the antibody signal. The data also shows that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.

References

This product has been referenced in:
  • Maurya VK  et al. Expression and activity of Rac1 is negatively affected in the dehydroepiandrosterone induced polycystic ovary of mouse. J Ovarian Res 7:32 (2014). Read more (PubMed: 24628852) »
  • Haas S  et al. DIGE proteome analysis reveals suitability of ischemic cardiac in vitro model for studying cellular response to acute ischemia and regeneration. PLoS One 7:e31669 (2012). WB ; Mouse . Read more (PubMed: 22384053) »

See all 4 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Rabbit Tissue lysate - whole (Brain)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
30 µg
Specification
Brain
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Aug 16 2017

The antibody concentration of the lot you have is 0.35 mg/ml.

This antibody recognizes a sequence that is exactly conserved amongst the proteins. We would thus expect that providing you resolve your gel in order to separate the two proteins, the antibody should recognise the two proteins separately.

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

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