Anti cbfβ

Manufactured by Santa Cruz Biotechnology

Anti-CBFβ is a laboratory reagent that can be used to detect and study the expression of the CBFβ protein in various biological samples. CBFβ is a subunit of the core binding factor (CBF) complex, which plays a crucial role in the regulation of gene expression. Anti-CBFβ is a specific antibody that can be used in techniques such as Western blotting, immunoprecipitation, and immunohistochemistry to analyze the presence, localization, and relative abundance of CBFβ in cells and tissues.

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Lab products found in correlation

Pierce supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (6 mentions) Lenalidomide, by Merck Group (2 mentions) Anti ikzf3, by Cell Signaling Technology (2 mentions) Anti ikzf1, by Cell Signaling Technology (2 mentions) Anti runx1, by Abcam (2 mentions) Anti crbn, by Merck Group (2 mentions) Anti ubiquitin k48, by Merck Group (2 mentions) Anti vinculin, by Santa Cruz Biotechnology (2 mentions) Anti rabbit igg hrp, by GE Healthcare (2 mentions) Anti mouse igg hrp, by GE Healthcare (2 mentions) Tnt t7 coupled reticulocyte lysate system, by Promega (2 mentions) Pomalidomide, by Merck Group (2 mentions) Anti tubulin, by Santa Cruz Biotechnology (2 mentions) Anti runx3, by Cell Signaling Technology (2 mentions) Anti flag, by Merck Group (2 mentions) Anti flag m2 affinity gel, by Merck Group (2 mentions) Glutathione sepharose 4b, by GE Healthcare (2 mentions) Benzonase, by Merck Group (2 mentions) Trans blot turbo transfer system, by Bio-Rad (2 mentions) Tgx gel, by Bio-Rad (1 mentions)

4 protocols using anti cbfβ

Characterizing CRBN-mediated Protein Degradation

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The following antibodies were used: anti-FLAG (Sigma, F7425), anti-IKZF3 (Cell Signaling, #15103), anti-IKZF1 (Cell Signaling, #9034), anti-RUNX1 (Abcam, ab92336), anti-RUNX3 (Cell Signaling, #9647), anti-CBFβ (Santa Cruz, sc-20693), anti-CRBN (Sigma, HPA045910), anti-ubiquitin (K48) (EMD Millipore, #05–1307), anti-TUBULIN (Santa Cruz, sc-8035), anti-VINCULIN (Santa Cruz, sc-73614), anti-Rabbit IgG-HRP (GE Healthcare, NA934V), and anti-mouse IgG-HRP (GE Healthcare, NA931V). The following agarose beads were used: anti-FLAG M2 Affinity Gel (Sigma, A2220) and Glutathione Sepharose 4B (GE Healthcare, #17075601). The following in vitro translation kit was used: TNT T7 Coupled Reticulocyte Lysate Systems (Promega, L4610). Benzonase was used according to manufacturer (Sigma, E1014). The following compounds were used: Lenalidomide (Sigma, CDS022536), Pomalidomide (Sigma, P0018), Bortezomib (Millennium Pharmaceuticals), and AI-10–104 was kindly provided by Dr. John H. Bushweller.

Zhou N., Gutierrez-Uzquiza A., Zheng X.Y., Chang R., Vogl D.T., Garfall A.L., Bernabei L., Saraf A., Florens L., Washburn M.P., Illendula A., Bushweller J.H, & Busino L. (2019). RUNX proteins desensitize multiple myeloma to lenalidomide via protecting IKZFs from degradation. Leukemia, 33(8), 2006-2021.

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Western Blot Analysis of Protein Complexes

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Whole cell extracts were run on 4–20% TGX gels (Bio-Rad) and blotted using the Trans-Blot Turbo transfer system (Bio-Rad) according to the manufacturer's instructions. Blots were blocked with 4% milk powder in 0.05% TBS-Tween and incubated with anti-HA (1:1000 dilution, H6908 Sigma), anti-CBFβ (1:400, sc20693 Santa Cruz), and anti-GAPDH antibodies (1:8000 dilution, ab8245 Abcam). Proteins were visualized using Pierce SuperSignal West Pico Chemiluminescent substrate (Thermo Scientific).

Illendula A., Gilmour J., Grembecka J., Tirumala V.S., Boulton A., Kuntimaddi A., Schmidt C., Wang L., Pulikkan J.A., Zong H., Parlak M., Kuscu C., Pickin A., Zhou Y., Gao Y., Mishra L., Adli M., Castilla L.H., Rajewski R.A., Janes K.A., Guzman M.L., Bonifer C, & Bushweller J.H. (2016). Small Molecule Inhibitor of CBFβ-RUNX Binding for RUNX Transcription Factor Driven Cancers. EBioMedicine, 8, 117-131.

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Immunoprecipitation Assay for E3 Ligase Complexes

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The following antibodies were used: anti-FLAG (Sigma, F7425), anti-IKZF3 (Cell Signaling, #15103), anti-IKZF1 (Cell Signaling, #9034), anti-RUNX1 (Abcam, ab92336), anti-RUNX3 (Cell Signaling, #9647), anti-CBFβ (Santa Cruz, sc-20693), anti-CRBN (Sigma, HPA045910), anti-ubiquitin (K48) (EMD Millipore, #05-1307), anti-TUBULIN (Santa Cruz, sc-8035), anti-VINCULIN (Santa Cruz, sc-73614), anti-Rabbit IgG-HRP (GE Healthcare, NA934V), and anti-mouse IgG-HRP (GE Healthcare, NA931V). The following agarose beads were used: anti-FLAG M2 Affinity Gel (Sigma, A2220) and Glutathione Sepharose 4B (GE Healthcare, #17075601). The following in vitro translation kit was used: TNT T7 Coupled Reticulocyte Lysate Systems (Promega, L4610). Benzonase was used according to manufacturer (Sigma, E1014). The following compounds were used: Lenalidomide (Sigma, CDS022536), Pomalidomide (Sigma, P0018), Bortezomib (Millennium Pharmaceuticals), and AI-10-104 was kindly provided by Dr. John H. Bushweller.

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Co-Immunoprecipitation and Western Blot Analysis

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Co-IPs were performed using the Pierce Crosslink Immunoprecipitation Kit (Thermo Fisher) according to manufacturer’s instructions. Samples were subsequently run on 4–20% gradient gels (Bio-Rad) and transferred to nitrocellulose membranes using a BioRad Trans-Blot Turbo transfer system. Membranes were blocked in 5% milk powder in 0.05% TBS-Tween and incubated with anti-HA (for RUNX1) (Sigma, H6908), anti-BRD4 (Bethyl Labs, A301-985A-100) and anti-CBFβ (Santa Cruz, sc20693) antibodies. Proteins were visualised using Pierce SuperSignal West Pico Chemiluminescent substrate (Thermo Scientific).
Western blots showing dose response of RUNX1 protein to Dox were performed as above then protein was detected using primary antibodies directed to AML1 (RUNX1) (Calbiochem, PC284) or GAPDH (Abcam, ab8245) followed by secondary antibodies IRDye® 800CW Donkey anti-Rabbit IgG (Li-Cor Biosciences) and IRDye® 800CW Donkey anti-Mouse IgG (Li-Cor Biosciences) respectively. Proteins were visualised on a Li-Cor Odyssey Imaging system.

Gilmour J., Assi S.A., Noailles L., Lichtinger M., Obier N, & Bonifer C. (2018). The Co-operation of RUNX1 with LDB1, CDK9 and BRD4 Drives Transcription Factor Complex Relocation During Haematopoietic Specification. Scientific Reports, 8, 10410.

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