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).
Ab76417
Manufactured by Abcam
Sourced in United States
Ab76417 is a laboratory product manufactured by Abcam. It serves as a tool for researchers to conduct various experiments and analyses. The core function of this product is to provide a means for researchers to achieve their experimental goals, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Ab134146, by Abcam (3 mentions)
Ripa lysis buffer, by Merck Group (2 mentions)
Ab54365, by Abcam (2 mentions)
Ab6258, by Abcam (2 mentions)
Ab6528, by Abcam (2 mentions)
Ab32358, by Abcam (2 mentions)
Ab51608, by Abcam (2 mentions)
Hrp conjugated goat anti rabbit igg h l, by Abcam (2 mentions)
Ab92547, by Abcam (2 mentions)
Ab32572, by Abcam (2 mentions)
Mini protean tgx stain free precast gel, by Bio-Rad (1 mentions)
Ripa buffer, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Total β catenin, by Cell Signaling Technology (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Phosphorylated β catenin, by Cell Signaling Technology (1 mentions)
Chemidoc chemiluminescence imaging system, by Bio-Rad (1 mentions)
Odyssey clx fluorescence imaging system, by LI COR (1 mentions)
Nonphosphorylated β catenin, by Cell Signaling Technology (1 mentions)
3 protocols using ab76417
1
Protein Expression Profiling in WiT49 Cells
2
Quantifying AML Cell Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AML cells (MV4-11 and MOLM-13) were drugged for 24 h, followed by protein extraction using RIPA buffer (Cell Signaling Technology) supplemented with PMSF. Cell pellets were incubated in RIPA buffer with constant agitation for 30 min, followed by sonication. Protein lysate was quantified using Pierce™ BCA Protein Assay Kit (ThermoFischer Scientific). 30 µg of protein was loaded into a Mini-PROTEAN TGX Stain-Free Precast Gel (Bio-Rad) and ran at 120 V for 1–2 h. Blots were developed using a ChemiDoc Chemiluminescence Imaging System (Bio-Rad) or by the Odyssey CLX Fluorescence Imaging System (Li-COR). Antibodies used in the presented studies purchased from Cell Signaling Technology included total β-catenin (#8480S), phosphorylated β-catenin (#9561S), non-phosphorylated β-catenin (cat#8814S), and β-actin (#3700S). Antibodies purchased from Abcam include LRP6 (ab134146) and phosphorlated-LRP6 (ab76417).
3
Protein Expression Analysis in WiT49 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!