Anti rac1 antibody

Manufactured by Abcam

Sourced in United Kingdom, United States

Anti-Rac1 antibody is a protein that specifically recognizes and binds to the Rac1 protein, a member of the Rho family of small GTPases. Rac1 is involved in various cellular processes, including cell migration, cell adhesion, and cytoskeletal organization. The Anti-Rac1 antibody can be used to detect and quantify the Rac1 protein in biological samples.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

RAC1 antibody Anti-Rac1

6 protocols using anti rac1 antibody

Rac1 Activation Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were washed in ice-cold PBS rapidly and lysed on ice in 50-mM Tris–HCl (pH 7.4), 1 % Triton X-100, 10 % glycerol, 2 mMMgCl₂, 100 mM NaCl, and protein inhibitor mixture. Lysates were centrifuged at 17,000×g at 4 °C for 5 min, and samples were taken from the supernatant to estimate total protein concentration. Twenty microgram of GST-Rac1 fusion bound to sepharose beads were added to cell lysate and incubated at 4 °C for 30 min. Beads were washed in lysis buffer four times, and bound proteins were eluted in Laemmli sample buffer. Total proteins and Rac1 affinity-purified proteins were analyzed by Western blotting using anti-Rac1 antibody (Abcam, Cambridge, UK).

Wu Y., Xu H., Li L., Yuan W., Zhang D, & Huang W. (2016). Susceptibility to Aspergillus Infections in Rats with Chronic Obstructive Pulmonary Disease via Deficiency Function of Alveolar Macrophages and Impaired Activation of TLR2. Inflammation, 39, 1310-1318.

+ Open protocol

+ Expand

Antibody-based Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies were all purchased from Cell Signaling Technology (CST, MA, USA): anti-JNK, anti-phospho-JNK, anti-c-JUN, anti-phospho-c-JUN, anti-Cyclin-D1, anti-AKT2, anti-phospho-AKT2, anti-cleaved PARP89, anti-mouse lgG HRP-linked antibody, anti-rabbit lgG HRP-linked antibody. β-actin, mouse mAb was purchased from YCASEN (YCASEN, Shanghai, China). Anti-Rac1 antibody, anti-MCL1 antibody and anti-PARP antibody [E51] were purchased from Abcam (Cambridge, MA, USA).

Zhao J., Jie Q., Li G., Li Y., Liu B., Li H., Luo J., Qin X., Li Z, & Wei Y. (2018). Rac1 promotes the survival of H9c2 cells during serum deficiency targeting JNK/c-JUN/Cyclin-D1 and AKT2/MCL1 pathways. International Journal of Medical Sciences, 15(10), 1062-1071.

+ Open protocol

+ Expand

Immunoblot Analysis of Rho GTPases

Check if the same lab product or an alternative is used in the 5 most similar protocols

All lysis buffers were supplemented with 1% protease inhibitor (Sigma Aldrich, USA). Protein extracts were resolved using gradient precast SDS – polyacrylamide gel electrophoresis (Biorad Laboratories Inc, USA), and then electro-transferred onto a nitrocellulose membrane by using Trans-Blot® Turbo™ Transfer System (Biorad Laboratories Inc, USA) for immunoblot analysis. Antibody probing was done as per manufacturers’ instructions Antibodies used include anti-RhoA antibody (1:1000, Abcam, UK), anti-CDC42 antibody (1:1000, Abcam, UK), anti-Rac1 antibody (1:1000 Abcam, UK), anti-phospho-MYPT1 (Thr696) antibody (1:1000, Cell Signaling Technology, USA), anti-myosin light chain (phospho S20) antibody (1:1000, Abcam, UK), anti-GAPDH antibody (1:2000, Abcam, UK). Secondary antibodies either IRDye® 800CW Goat anti-Mouse IgG (LI-COR, USA) or IRDye® 800CW Goat anti-Rabbit IgG (LI-COR, USA) were used with dilution of 1:15000. Specific protein bands were detected using Odyssey® CLx Imaging System (LI-COR, USA).

Xu J., Yang J., Nyga A., Ehteramyan M., Moraga A., Wu Y., Zeng L., Knight M.M, & Shelton J.C. (2018). Cobalt (II) ions and nanoparticles induce macrophage retention by ROS-mediated down-regulation of RhoA expression. Acta Biomaterialia, 72, 434-446.

+ Open protocol

+ Expand

Rac1 Activation Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Rac1 activation assays were conducted as previously described to test the activation of Rac1 in cells (22 (link)). Briefly, cells were rinsed with cold PBS and lysed in ice-cold radioimmunoprecipitation assay (RIPA) buffer. Lysates were cleared by centrifugation and the protein concentration in the clarified supernatant was measured. Equal amounts of protein (400–600 µg) were incubated with 50 µg of PAK1 agarose beads (Assay Designs, Farmingdale, NY, USA), and the reaction mixture was gently rocked at 4°C for 60 min. The agarose beads were collected by pulsing for 5 sec in a microcentrifuge at 14,000 × g and the supernatant was removed and discarded. The beads were washed thrice with lysis buffer, and the agarose beads were resuspended in 40 µl of 2× Laemmli buffer and boiled for 10 min to elute the bound proteins. Eluted proteins were resolved in 12% polyacrylamide gels, transferred to polyvinylidene difluoride (PVDF) membranes, and immunoblotted with the anti-Rac1 antibody (Abcam, Cambridge, UK).

Zhao K., Wang L., Li T., Zhu M., Zhang C., Chen L., Zhao P., Zhou H., Yu S, & Yang X. (2017). The role of miR-451 in the switching between proliferation and migration in malignant glioma cells: AMPK signaling, mTOR modulation and Rac1 activation required. International Journal of Oncology, 50(6), 1989-1999.

+ Open protocol

+ Expand

Quantifying Active Rac1 in Brain Lysates

Check if the same lab product or an alternative is used in the 5 most similar protocols

The activation form of Rac1 (GTP-Rac1) was obtained by a pull-down assay using Rac1 activation Assay kit (Abcam, USA). According to the instructions, brain lysates were incubated with PAK1 PBD bead at 4°C for 1 h. The eluted proteins were then detected by performing Western blotting with anti-Rac1 antibody (Abcam, USA) as described above.

Liu W., Huang J., Doycheva D., Gamdzyk M., Tang J, & Zhang J.H. (2019). RvD1binding with FPR2 attenuates inflammation via Rac1/NOX2 pathway after neonatal hypoxic-ischemic injury in rats. Experimental neurology, 320, 112982.

+ Open protocol

+ Expand

Rac1 and GAPDH Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot analysis was carried out using standard methods. Proteins were separated on 10% SDS-PAGE, and then transferred to PVDF membranes (Amersham, Buckinghamshire, UK). Membranes were blocked overnight with 5% non-fat dried milk and incubated for 2 h with anti-Rac1 antibody (Abcam, England) at 1∶1000 dilution; anti-GAPDH antibody (Proteintech,Chicago, USA) at 1∶50,000 dilution. After washing with TBST (10 mM Tris, pH 8.0, 150 mM NaCl, and 0.1% Tween20), the membranes were incubated for 2 h with goat anti-rabbit antibody (zsgb-bio, Beijing, China) at 1∶5000 dilution and 1∶50000 dilution.

Geng S., Zhang X., Chen J., Liu X., Zhang H., Xu X., Ma Y., Li B., Zhang Y., Bi Z, & Yang C. (2014). The Tumor Suppressor Role of miR-124 in Osteosarcoma. PLoS ONE, 9(6), e91566.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!