Cul4a

Manufactured by Fortis Life Sciences

Cul4A is a laboratory equipment product. It serves as a core component for specific scientific applications. The detailed technical specifications and intended use of this product are not available at this time.

Automatically generated - may contain errors

Lab products found in correlation

3 protocols using cul4a

Comprehensive Antibody Resource for Cell Biology

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

Anti-PAR6β (BC31AP), anti-Lgl2 (N13AP), and anti-Lgl2-S653P antibodies have been described previously (Yamanaka et al., 2003 (link)). Anti-GP135 (3F2/D8) was a kind gift from George K. Ojakian (State University of New York, Brooklyn, NY). Other antibodies were purchased as follows: Lgl2 (Abnova, Taipei, Taiwan), Lgl1 (Sigma-Aldrich, St. Louis, MO), VprBP (Proteintech Group, Chicago, IL), DDB1 (Bethyl, Montgomery, TX), Cul4A (Bethyl), Cul4B (Proteintech Group), Cdt2 (Novus Biologicals, Littleton, CO), PKC iota (BD, Franklin Lakes, NJ), zonula occludens-1 (ZO-1; Santa Cruz Biotechnology, Dallas, TX), p27kip1 (BD), p16INK4a (Cell Signaling, Danvers, MA), p21Waf1/Cip1 (Santa Cruz Biotechnology), Skp2 (Santa Cruz Biotechnology), Cdh1 (Abcam, Cambridge, United Kingdom), cyclin A (Santa Cruz Biotechnology), cyclin B1 (Santa Cruz Biotechnology), HSP70 (Enzo Life Sciences, Farmingdale NY), EDD1 (Bethyl), glyceraldehyde-3-phosphate dehydrogenase (Abcam), β-Actin (Sigma-Aldrich), E-cadherin (Sigma-Aldrich), V5 (Invitrogen), HA (Roche, Basel, Switzerland), SBP (Santa Cruz Biotechnology), Myc (Millipore, Billerica, MA; Cell Signaling), BrdU (BD; Abcam), and normal rabbit immunoglobulin G (Cell Signaling).

Yamashita K., Ide M., Furukawa K.T., Suzuki A., Hirano H, & Ohno S. (2015). Tumor suppressor protein Lgl mediates G1 cell cycle arrest at high cell density by forming an Lgl-VprBP-DDB1 complex. Molecular Biology of the Cell, 26(13), 2426-2438.

+ Open protocol

+ Expand

Immunoblotting of DNA Damage Signaling

Cited 1 time

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

Cell pellets were harvested and then lysed in a buffer consisting of: 10mM HEPES pH 7.5, 10mM KCl, 1% NP-40, with phosphatase inhibitor (PhosSTOP from Sigma Aldrich) and a protease inhibitor cocktail (Roche cOmplete EDTA-free). Lysates were cleared of chromatin and membranous debris by centrifugation (10–15 minutes at 21,100 x g) and lysate concentrations were determined by Bradford (Biorad) or BCA assay (Pierce).
A total of 30–60 μgs of protein was loaded per lane. Proteins were separated on a 4–12% Bis-Tris gradient gel (Biorad) and blotted onto PVDF (Millipore) or nitrocellulose (GE).
Antibodies utilized are as follows: HUWE1/Lasu1/Ureb1 (Bethyl), Chk1 (SCBT), pATM S1981 (CST), ATM (CST), pATR T1989 (CST), ATR (CST), pChk1 S296 (CST), Actin-HRP (Abcam), GFP (GeneTex), MDM2 (R&D Systems), Cul4A (Bethyl), HA (SCBT), p53 (SCBT), γ-H2AX (CST) and H2AX (CST). Blots were incubated overnight in primary antibody at 4°C with the exception of HRP-conjugated Actin, which was incubated for 1 hour at room temperature.
Immunoblots were quantified using ImageJ and values were plotted in Excel.
The Chk1-eGFP expression construct was generously provided by Dr. Youwei Zhang (Case Western Reserve University, Cleveland, Ohio). The HA-Ub expression construct was described previously [49 (link)].

Cassidy K.B., Bang S., Kurokawa M, & Gerber S.A. (2019). Direct regulation of Chk1 protein stability by E3 ubiquitin ligase HUWE1.. The FEBS journal, 287(10), 1985-1999.

+ Open protocol

+ Expand

Melanoma Protein Signaling Profiling

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

The melanoma tumor tissues and cultured cells were harvested in Tween 20 buffer containing 50mM HEPES (pH 8.0), 150mM NaCl, 2.5mM EGTA, 1mM EDTA, 0.1% Tween 20, and protease/phosphatase inhibitors (1mM phenylmethylsulphonyl fluoride, 20 U of aprotinin/ml, 5mg of leupeptin/ml, 1mM DTT, 0.4mM NaF, and 10mM β-glycerophosphate). Lysates were sonicated prior to clearing by centrifugation at 4°C for 30 min. Proteins were resolved by SDS-PAGE, transferred to membrane, and subjected to immunoblot. Antibodies utilized include PERK (Rockland), p-eIF2α S51 (Cell Signaling), BiP (Cell Signaling), total eIF2α (Cell Signaling), Cyclin D1 (mouse monoclonal D1-72-13G), Cul4a (Bethyl, A300-739A), p-AktS473 (Cell Signaling), total Akt (Cell Signaling), GAPDH (Cell Signaling) and β-actin (Sigma Aldrich).

Pytel D., Gao Y., Mackiewicz K., Katlinskaya Y.V., Staschke K.A., Paredes M.C., Yoshida A., Qie S., Zhang G., Chajewski O.S., Wu L., Majsterek I., Herlyn M., Fuchs S.Y, & Diehl J.A. (2016). PERK Is a Haploinsufficient Tumor Suppressor: Gene Dose Determines Tumor-Suppressive Versus Tumor Promoting Properties of PERK in Melanoma. PLoS Genetics, 12(12), e1006518.

+ 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!