Unless indicated, primary antibodies were used at a dilution of 1:1,000 for Western blotting. Anti-GAPDH was purchased from EMD Millipore and used at a dilution of 1:20,000. Rabbit anti-PAK1, rabbit anti–phospho-PAK4 (S474)/PAK5 (S602)/PAK6 (S560), and rabbit anti-PAK4, which also recognizes PAK6 (Ahmed et al., 2008 (link)), were purchased from Cell Signaling Technology. Rabbit polyclonal PAK4-specific antibody (raised against PAK4 peptide sequence CRRAGPEKRPKSSREG) has been described elsewhere (Wells et al., 2010 (link)). Rabbit anti-RhoU (Wrch1) and rabbit anti-Rab40A were obtained from Abcam and used at a dilution of 1:500. Mouse anti-GFP was obtained from Roche and mouse anti-T7 from EMD Millipore. Rabbit anti-HA (Y-11) and mouse c-Myc (9E10) were purchased from Santa Cruz Biotechnology, Inc. Mouse antipaxillin were obtained from BD, rabbit anti-phosphopaxillin S273(272) from Invitrogen, and mouse antivinculin from Sigma-Aldrich. HRP-conjugated secondary antibodies were purchased from Dako and diluted 1:2,000. Staurosporine was purchased from Cell Signaling Technology and dissolved in DMSO.
Mouse c myc 9e10
Manufactured by Santa Cruz Biotechnology
Mouse c-Myc (9E10) is a monoclonal antibody that specifically recognizes the c-Myc protein, a transcription factor that plays a crucial role in cell growth and proliferation. This antibody can be used for the detection and study of c-Myc in various applications, such as Western blotting, immunoprecipitation, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti gfp, by Roche (2 mentions)
Rabbit anti pak1, by Cell Signaling Technology (2 mentions)
Rabbit anti pak4, by Cell Signaling Technology (2 mentions)
Rabbit anti ha y11, by Santa Cruz Biotechnology (2 mentions)
Hrp conjugated secondary antibody, by Agilent Technologies (2 mentions)
Goat rat alexa488, by Thermo Fisher Scientific (2 mentions)
Anti gapdh, by Merck Group (2 mentions)
Staurosporine, by Cell Signaling Technology (1 mentions)
Mouse anti t7, by Merck Group (1 mentions)
Rabbit anti rhou wrch1, by Abcam (1 mentions)
Rabbit anti phosphopaxillin s273 272, by Thermo Fisher Scientific (1 mentions)
Mouse anti vinculin, by Merck Group (1 mentions)
Mouse anti paxillin, by BD (1 mentions)
4 protocols using mouse c myc 9e10
1
Detailed Antibody Dilutions and Reagent Use
2
Antibody Validation for Western Blotting
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Unless indicated, primary antibodies were used at a dilution of 1:1000 for Western blotting. Anti-GAPDH was purchased from Millipore and used at a dilution of 1:20000. Rabbit anti-PAK1, rabbit anti-phospho-PAK4 (Ser 474)/PAK5 (Ser 602)/ PAK6 (Ser 560) and rabbit anti-PAK4, which also recognizes PAK635 (link) were purchased from Cell Signaling Technology. Rabbit polyclonal PAK4 specific antibody (raised against PAK4 peptide sequence CRRAGPEKRPKSSREG) has been described elsewhere37 (link). Mouse anti-GFP was obtained from Roche. Rabbit anti-HA (Y-11) and mouse c-Myc (9E10) were purchased from Santa Cruz. HRP-conjugated secondary antibodies were purchased from DAKO and diluted 1:2000.
3
Immunostaining of Dechorionated Embryos
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Dechorionated embryos were fixed overnight at 4 °C in PEM (80 mM Sodium-Pipes, 5 mM EGTA, 1 mM MgCl2) - 4% PFA - 0.04% TritonX100. After washing 2 × 5 min in PEMT (PEM - 0.2% TritonX100), 10 min in PEM - 50 mM NH4Cl, 2 × 5 min in PEMT and blocking in PEMT - 5% NGS, embryos were incubated 2 hrs at room temperature with primary antibodies. Following incubation, embryos were washed during 5, 10, 15 and 20 min in PEMT, blocked in PEMT - 5% NGS, and incubated again with secondary antibodies for 2 hrs. Embryos were again washed during 5, 10, 15, and 20 min in PEMT. The following primary antibodies were used: Rat@HA (Roche 11 867 423 001, 1:500). Mouse@c-Myc9E10 (Santa Cruz sc-40, 1:500). Secondary antibodies Goat@Rat-Alexa488 (Invitrogen) and Goat@Mouse-Cy5 (Jackson Immunoresearch) were used at a dilution of 1:500.
4
Immunofluorescence Analysis of Dechorionated Embryos
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Dechorionated embryos were fixed overnight at 4°C in PEM (80 mM Sodium-Pipes, 5 mM EGTA, 1 mM MgCl2) -4% PFA -0.04% TritonX100. After washing 2 x 5 min in PEMT (PEM -0.2% TritonX100), 10 min in PEM -50 mM NH4Cl, 2 x 5 min in PEMT and blocking in PEMT -5% NGS, embryos were incubated 2 hrs at room temperature with primary antibodies. Following incubation, embryos were washed during 5, 10, 15 and 20 min in PEMT, blocked in PEMT -5% NGS, and incubated again with secondary antibodies for 2 hrs.
Embryos were again washed during 5, 10, 15 and 20 min in PEMT. The following primary antibodies were used: Rat@HA (Roche 11 867 423 001, 1:500). Mouse@c-Myc9E10 (Santa Cruz sc-40, 1:250). Secondary antibodies Goat@Rat-Alexa488 (Invitrogen) and Goat@Mouse-Cy5 (Jackson Immunoresearch) were used at a dilution of 1:500.
Embryos were again washed during 5, 10, 15 and 20 min in PEMT. The following primary antibodies were used: Rat@HA (Roche 11 867 423 001, 1:500). Mouse@c-Myc9E10 (Santa Cruz sc-40, 1:250). Secondary antibodies Goat@Rat-Alexa488 (Invitrogen) and Goat@Mouse-Cy5 (Jackson Immunoresearch) were used at a dilution of 1:500.
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