Ras antibody

Manufactured by Cell Signaling Technology

Sourced in United States

The RAS antibody is a research tool designed to detect and quantify RAS proteins, which are small GTPases involved in cell signaling pathways. This antibody can be used to identify and study the expression of RAS proteins in various biological samples.

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Lab products found in correlation

Amylose resin, by Thermo Fisher Scientific (3 mentions) Glutathione sepharose 4b resin, by Cytiva (3 mentions) Pierce streptavidin magnetic beads, by Thermo Fisher Scientific (3 mentions) Sc 138, by Cell Signaling Technology (3 mentions) Typhoon imager, by Cytiva (3 mentions) Phospho stat6, by Cell Signaling Technology (1 mentions) β actin antibody, by Cell Signaling Technology (1 mentions) Jak1 antibody, by Cell Signaling Technology (1 mentions) Phospho stat5, by Cell Signaling Technology (1 mentions) Mmp 9 antibody, by Cell Signaling Technology (1 mentions) Phospho jak2, by Cell Signaling Technology (1 mentions) Pkcζ antibody, by Cell Signaling Technology (1 mentions) Plcγ1 antibody, by Cell Signaling Technology (1 mentions) Phospho stat2, by Cell Signaling Technology (1 mentions) Phospho stat1, by Cell Signaling Technology (1 mentions) Phospho stat3, by Cell Signaling Technology (1 mentions) Gapdh mouse monoclonal antibody, by Proteintech (1 mentions) Phospho akt ser473 d9e xp rabbit mab, by Cell Signaling Technology (1 mentions) Sds page buffer, by Beyotime (1 mentions) Tgrinder, by Tiangen Biotech (1 mentions)

Spelling variants of this product

Anti-Ras antibody (4 citations) K-Ras (G12D) antibody (2 citations)

5 protocols using ras antibody

Affinity Capture of HRAS, KRAS, and BRAF Proteins

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All proteins were added in 1:1 stoichiometric ratio and incubated together in binding buffer (50 mM HEPES pH 7.4, 150 mM NaCl, 5% glycerol, and 0.125 mg/mL BSA) with 20 µL of Glutathione Sepharose 4b resin (Cytiva), amylose resin (NEB), or Pierce streptavidin magnetic beads (Thermo Scientific). After extensive washing (50 mM HEPES pH 7.4, 500 mM NaCl, 5% glycerol, and 0.125 mg/mL BSA), 30 µL 4× loading dye was added to resin. Supernatants were loaded and protein analyzed through SDS-PAGE. After transfer to nitrocellulose membrane, GST-HRAS/-KRAS was probed with GST antibody (Santa Cruz Biotechnologies SC-138), KRAS with RAS antibody (Cell Signaling 67648S), and BRAF NTs/ BRAF-KD with His antibody (Sigma SAB5600227). Finally, western blots were imaged on Cytiva Typhoon imager.

Trebino T.E., Markusic B., Nan H., Banerjee S, & Wang Z. (2023). Unveiling the domain-specific and RAS isoform-specific details of BRAF kinase regulation. eLife, 12, RP88836.

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Protein Kinase Signaling Pathway Analysis

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The following antibodies were purchased from Cell Signaling (Danvers, MA, USA): JAK1 Antibody (#3332, 130kDa), Phospho-JAK2 (Tyr1007/1008) Antibody (#3771, 125kDa), β-Actin Antibody (#4967, 45kDa), Phospho-STAT1 (Tyr701) (58D6) Rabbit mAb (#9167, 84,91kDa), Phospho-STAT2 (Tyr690) Antibody (#4441, 113kDa), Phospho-STAT3 (Tyr705) Antibody (#9131, 79,86kDa), Phospho-STAT5 (Tyr694) Antibody (#9351, 90kDa), Phospho-STAT6 (Tyr641) Antibody (#9361, 110kDa), Ras Antibody (#3965, 21kDa), p44/42-MAPK (Erk1/2) Antibody (#9102, 42,44kDa), GSK-3β (27C10) Rabbit mAb (#9315, 46kDa), MMP-9 Antibody (#3852, 84,92kDa), Phospho-PKCα/β II (Thr638/641) Antibody (#9375, 80,82kDa); PKCα Antibody (#2056, 80kDa); Phospho-PKC (pan) (βII Ser660) Antibody (#9371, 78 a 85kDa); Phospho-PKD/PKCμ (Ser744/748) Antibody (#2054, 115kDa); PKD/PKCμ Antibody (#2052, 115kDa); PKCδ Antibody (#2058, 78kDa); Phospho-PKCδ/θ (Ser643/676) Antibody (#9376, 78kDa), PKCζ Antibody (#9372, 78kDa); PLCγ1 Antibody (#2822, 155kDa); anti-mouse, anti-rabbit and anti-goat IgGs antibodies. From Abcam (Cambridge, MA, USA): anti-PKC antibody (ab59363) was purchased. PepChip1-Kinomics^™ slides were obtained from Pepscan Presto BV (Lelystad, the Netherlands).
WP1066 (#573097; C₁₇H₁₄BrN₃O) and Tofacitinib (#PZ0017; C₁₆H₂₀N₆O · C₆H₈O₇) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Nascimento A.S., Peres L.L., Fari A.V., Milani R., Silva R.A., da Costa Fernandes C.J., Peppelenbosch M.P., Ferreira-Halder C.V, & Zambuzzi W.F. (2017). Phosphoproteome profiling reveals critical role of JAK-STAT signaling in maintaining chemoresistance in breast cancer. Oncotarget, 8(70), 114756-114768.

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Protein-Protein Interaction Assay

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All proteins were added in 1:1 stoichiometric ratio and incubated together in binding buffer (50 mM HEPES pH 7.4, 150 mM NaCl, 5% Glycerol, and 0.125 mg/mL BSA) with 20 μL of glutathione sepharose 4b resin (Cytiva), amylose resin (NEB), or Pierce streptavidin magnetic beads (ThermoScientific). After extensive washing (50 mM HEPES pH 7.4, 500 mM NaCl, 5% Glycerol, and 0.125 mg/mL BSA), 30 μL 4x loading dye was added to resin. Supernatant were loaded and protein analyzed through SDS-PAGE. After transfer to nitrocellulose membrane, GST-HRAS/-KRAS was probed with GST antibody (Santa Cruz Biotechnologies SC-138), KRAS with RAS antibody (Cell Signaling 67648S) and BRAF NTs/ BRAF KD with His antibody (Sigma SAB5600227). Finally, western blots were imaged on Cytiva Typhoon imager.

Trebino T., Markusic B., Nan H., Banerjee S, & Wang Z. (2023). Unveiling the Domain-Specific and RAS Isoform-Specific Details of BRAF Regulation. bioRxiv.

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Cellular Signaling Pathway Analysis

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A549 and H2228 cells were plated in six-well plates at a confluency of 70%. 48 hours after adenovirus infection, whole-cell extracts were prepared by lysing cells with the addition of 500 µL of hot SDS-PAGE buffer (Beyotime, P0015B). Tumor tissues were homogenized by TGrinder (Tiangen, OSE-Y30), and lysed with RIPA buffer containing complete protease inhibitor cocktail (Roche, 11697498001). Target proteins were detected by western blot analysis with the following antibodies: GAPDH mouse monoclonal antibody (Proteintech, 60004-1-Ig,), Akt (pan) (40D4) mouse monoclonal antibody (Cell Signaling Technology, 2920), Phospho-Akt (Ser473) (D9E) XP Rabbit mAb (Cell Signaling Technology, 4060), p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb (Cell Signaling Technology, 4695), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) (Cell Signaling Technology, 4370), mouse monoclonal Anti-MAP Kinase, activated (Diphosphorylated ERK-1&2) antibody (Sigma-Aldrich, M8159), Ras Antibody (Cell Signaling Technology, 3965), and Anti-RAS (G12S) Mouse Monoclonal Antibody (NewEast Biosciences, 26186).

Gao Q., Ouyang W., Kang B., Han X., Xiong Y., Ding R., Li Y., Wang F., Huang L., Chen L., Wang D., Dong X., Zhang Z., Li Y., Ze B., Hou Y., Yang H., Ma Y., Gu Y, & Chao C.C. (2020). Selective targeting of the oncogenic KRAS G12S mutant allele by CRISPR/Cas9 induces efficient tumor regression. Theranostics, 10(11), 5137-5153.

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Protein-Protein Interaction Assay

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All proteins were added in 1:1 stoichiometric ratio and incubated together in binding buffer (50 mM HEPES pH 7.4, 150 mM NaCl, 5% Glycerol, and 0.125 mg/mL BSA) with 20 μL of glutathione sepharose 4b resin (Cytiva), amylose resin (NEB), or Pierce streptavidin magnetic beads (ThermoScientific). After extensive washing (50 mM HEPES pH 7.4, 500 mM NaCl, 5% Glycerol, and 0.125 mg/mL BSA), 30 μL 4x loading dye was added to resin. Supernatant were loaded and protein analyzed through sds-PAGE. After transfer to nitrocellulose membrane, GST-HRAS/-KRAS was probed with GST antibody (Santa Cruz Biotechnologies SC-138), KRAS with RAS antibody (Cell Signaling 67648S) and BRAF NTs/ BRAF KD with His antibody (Sigma SAB5600227). Finally, western blots were imaged on Cytiva Typhoon imager.

Trebino T., Markusic B., Nan H., Banerjee S, & Wang Z. (2023). Unveiling the Domain-Specific and RAS Isoform-Specific Details of BRAF Regulation. bioRxiv.

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