The levels of HOXA11, β-catenin, receptor–related protein 6 (LRP6), phosphorylation-LRP6 (p-LRP6), E-cadherin (E-cad), N-cadherin (N-cad), Vimentin, HIF-1α, C/EBPβ, and β-actin protein expression were assessed in WiT49 cells. The total cellular proteins were isolated with RIPA lysis buffer (#R0278, Sigma), and a standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method was used to separate the proteins [19 ]. Next, the separated protein bands were transferred onto membranes that were subsequently blocked, and then cultured with primary antibodies against HOXA11 (#ab54365; 1:1000), β-catenin (#ab32572; 1:800), LRP6 (#ab134146; 1:1000), p-LRP6 (#ab76417; 1:800), E-cad (#ab6528; 1:800), N-cad (#ab6258; 1:800), Vimentin (#ab92547; 1:800), HIF-1α (#ab51608; 1:1000), C/EBPβ (#ab32358; 1:1000), and β-actin (#ab32572; 1:5000) (Abcam, Cambridge, MA, USA). Next, the membranes were then washed and incubated with HRP conjugated Goat Anti-Rabbit IgG H&L (1: 5000, #ab6721, Abcam). Finally, the immunostained bands were observed using a Tanon 6600 Luminescence imaging workstation (Tanon, China).
Ab6258
Manufactured by Abcam
Sourced in United States
Ab6258 is a monoclonal antibody that recognizes the human Protein X. It is intended for use in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab6528, by Abcam (2 mentions)
Ab134146, by Abcam (2 mentions)
Ab32358, by Abcam (2 mentions)
Ab51608, by Abcam (2 mentions)
Ab54365, by Abcam (2 mentions)
Ab76417, by Abcam (2 mentions)
Ripa lysis buffer, by Merck Group (2 mentions)
Hrp conjugated goat anti rabbit igg h l, by Abcam (2 mentions)
Ab92547, by Abcam (2 mentions)
Ab32572, by Abcam (2 mentions)
2 protocols using ab6258
1
Protein Expression Profiling in WiT49 Cells
2
Protein Expression Analysis in WiT49 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The protein expression levels of HOXA11, β-catenin, receptor-related protein 6 (LRP6), Phosphorylation-LRP6 (p-LRP6), E-cadherin (E-cad), N-cadherin (N-cad), Vimentin, HIF-1α, C/EBPβ, and β-actin were assessed in WiT49 cells. The proteins were isolated by RIPA lysis buffer (#R0278, Sigma). Standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method was used to separate the proteins [20] . Then, the gels were transferred, blocked, and cultured with primary antibodies against HOXA11 (#ab54365; 1:1000), β-catenin (#ab32572; 1:800), LRP6 (#ab134146; 1:1000), p-LRP6 (#ab76417; 1:800), E-cad (#ab6528; 1:800), N-cad (#ab6258; 1:800), Vimentin (#ab92547; 1:800), HIF-1α (#ab51608; 1:1000), C/EBPβ (#ab32358; 1:1000), and β-actin (#ab32572; 1:5000) bought from Abcam (Cambridge, MA, USA). The membranes were washed and incubated with the HRP conjugated Goat Anti-Rabbit IgG H&L (1: 5000, #ab6721, Abcam). Finally, the bands were observed under a Tanon 6600 Luminescence imaging workstation (Tanon, China). Bioinformatics analysis PROMO (http://alggen.lsi.upc.es/cgi-bin/promo_v3/promo/promoinit.cgi?dirDB=TF_8.3) and JASPAR(http://jaspar.genereg.net/) databases were used for the transcription factor prediction of HOXA11-AS with the threshold.
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