Anti bim c34c5

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-Bim (C34C5) is a mouse monoclonal antibody that recognizes the Bim protein. Bim is a pro-apoptotic member of the Bcl-2 family and plays a role in the intrinsic apoptosis pathway.

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4 protocols using anti bim c34c5

Targeted Cancer Therapy Compounds and Antibodies

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The following drugs were purchased: Dinaciclib (SCH727965) for in vitro and in vivo studies (S2768; Selleckchem), lapatinib ditosylate (Tykerb) for in vitro and in vivo studies (M1802; Abmole), neratinib for in vivo studies (M1913; Abmole), A-1210477 (CT-A121; Chemietek), A-1331852 (22963; Cayman Chemicals), tucatinib (HY-16069; Medchem), and ABT-199 (venetoclax) (CT-A199; Chemietek). The antibodies used in this study were as follows: anti-Bak (AB-1 clone for IP) (AM03; EMD Millipore), anti-Bak (3814S; Cell Signaling), anti-Bim (C34C5) (2933S; Cell Signaling), anti–BCL-xL (54H6) (2764S; Cell Signaling), anti–Bcl-2 (D55G8) (Human Specific) (4223S; Cell Signaling), anti-cleaved PARP (Asp214) (D64E10) (5625S; Cell Signaling), anti-GAPDH (6C5) (sc-32233; Santa Cruz), anti–MCL-1 (S-19) (sc-819; Santa Cruz), anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) (4370S; Cell Signaling), anti-phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) (5364S; Cell Signaling), anti-phospho-Akt (Thr308) (244F9) (4056S; Cell Signaling), anti-phospho-Akt (Ser473) (D9E) (4060S; Cell Signaling), anti-HER2/ErbB2 (29D8) (2165S; Cell Signaling), anti-phospho-HER2/ErbB2 (Tyr1248) (2247S; Cell Signaling), anti-phospho-Rpb1 CTD (Ser 2/5) (4375S; Cell Signaling), Normal Rabbit IgG for IP (sc-2027; Santa Cruz), and Normal Mouse IgG for IP (sc-2025; Santa Cruz).

Floros K.V., Jacob S., Kurupi R., Fairchild C.K., Hu B., Puchalapalli M., E. Koblinski J., Dozmorov M.G., Boikos S.A., Scaltriti M, & Faber A.C. (2021). Targeting transcription of MCL-1 sensitizes HER2-amplified breast cancers to HER2 inhibitors. Cell Death & Disease, 12(2), 179.

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Investigating Cell Cycle Regulation

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Antibodies used for this study include rabbit polyclonal anti-CDC20 (H-175) (Santa Cruz Biotechnology), rabbit monoclonal anti-Bim (C34C5) (Cell Signaling Technology), rabbit polyclonal anti-p21 antibody (ab227443) (Abcam), mouse monoclonal anti-α-tubulin antibody (Sigma), and rabbit monoclonal anti-phospho-histone H2A.X (Ser139) (Cell Signaling Technology). Temozolomide was purchased from Tocris Bioscience.

Mao D.D., Cleary R.T., Gujar A., Mahlokozera T, & Kim A.H. (2022). CDC20 regulates sensitivity to chemotherapy and radiation in glioblastoma stem cells. PLoS ONE, 17(6), e0270251.

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Comprehensive Immune Cell Profiling

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Peripheral lymph nodes and/or spleens were harvested from mice with indicated genotypes, and grinded into single cells, subjected to followed experiments. For analysis of surface markers, cells were washed in PBS containing 2% (wt/vol) FBS, and stained with indicated Abs (dilution 1: 200); intracellular markers were strained with Transcription Factor Buffer Set (BD Pharmingen, 562574) according to the manufacturer’s instructions by indicated Abs (dilution 1: 50). For intracellular cytokine staining, cells were stimulated for 5 h with leukocyte activation cocktail of GolgiPlug (BD Pharmingen, 550583). Antibodies were all from eBioscience, unless otherwise noted: anti-CD4 (GK1.5), anti-CD8α (53-6.7, BD Pharmingen), anti-CD44 (IM7, BD Pharmingen), anti-CD62L (MEL-14), anti-Foxp3 (FJK-16s), anti-Ki67 (SolA15), anti-CD45 (30-F11), anti-F4/80 (BM8), anti-CD11b (M1/70), anti-CD11c (N418), anti-MHCII (M5/114.15.2), anti-CD80 (16-10A1), anti-CD86 (GL1, BD Pharmingen), anti-B220 (RA3-6B2), anti-CD19 (1D3, BD Pharmingen), anti-GL7 (GL7, BD Pharmingen), anti-Fas (15A7), anti-active caspase 3 (C92-605, BD Pharmingen), anti-Ifn-γ (XMG1.2), anti-Il4 (11B11), anti-Il17A (eBio17B7), and anti-Bim (C34C5, Cell Signal Technology). Flow cytometry data were acquired on CytoFLEX LX (Beckman) and analyzed using Flowjo software (Tree Star).

Wu D., Li H., Liu M., Qin J, & Sun Y. (2022). The Ube2m-Rbx1 neddylation-Cullin-RING-Ligase proteins are essential for the maintenance of Regulatory T cell fitness. Nature Communications, 13, 3021.

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Western Blot Analysis of Protein Signaling

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Cellular proteins were extracted using Laemmli buffer and protein concentration was measured using Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). Cell lysates were resolved by SDS-PAGE and transferred to 0.2 µm nitrocellulose Trans-Blot Turbo TM membranes (Thermo Fisher Scientific). For protein detection, the following antibodies were used according to the protocols supplied by the manufacturers: anti-MET phospho-Tyr1234/1235 (D26, Cell Signaling Technology); anti-MET (D1C2, Cell Signaling Technology); anti-AKT (Cell Signaling Technology); anti-AKT phospho Ser473 (Cell Signaling Technology); anti-ERK1/2 (Cell Signaling Technology); ERK1/2 phospho Thr202/Tyr204 (Cell Signaling Technology); anti-vinculin (clone hVIN-1, Sigma-Aldrich); anti-BIM (C34C5, Cell Signaling Technology); anti-BCL-xL (54H6, Cell Signaling Technology); anti-BCL2 (M19, Santa Cruz Biotechnology Inc., Dallas, TX, USA); anti-alpha actin (Sigma-Aldrich); anti-caspase-3 antibody that detects full length and cleaved caspase-3 forms (Cell Signaling Technology); anti-tubulin (Sigma-Aldrich). HRP-conjugated secondary antibodies were from Jackson Immuno Research Europe (Cambridge, UK). Protein detection and quantification were performed using the ChemiDoc Touch Imaging System (Bio-Rad, Hercules, CA, USA) and the Image Lab software (Bio-Rad).

Modica C., Gallo S., Chiriaco C., Spilinga M., Comoglio P.M., Crepaldi T., Basilico C, & Vigna E. (2020). Molecular Engineering Strategies Tailoring the Apoptotic Response to a MET Therapeutic Antibody. Cancers, 12(3), 741.

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