Rabbit anti bak

Manufactured by Cell Signaling Technology

Sourced in United Kingdom

Rabbit anti-BAK is a primary antibody that specifically recognizes the pro-apoptotic protein BAK. BAK is a member of the Bcl-2 family of proteins and plays a key role in the intrinsic apoptotic pathway. This antibody can be used to detect and study the expression and localization of BAK in various experimental systems.

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5 protocols using rabbit anti bak

Antibody Profiling for Apoptosis Signaling

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Antibodies used for immunoblotting analyses in this study are listed below: rabbit anti-MCL-1 (1:500, ab32087), rabbit anti-BAX (1:1000, ab32503) were obtained from Abcam; mouse anti-BCL-2 (1:1000, #15071), rabbit anti-BIM (1:1000, #2933), rabbit anti-BCL-XL (1:1000, #2762), rabbit anti-BAK (1:1000, #12105), rabbit anti-AKT (1:1000, #4691), rabbit anti-p-AKT (1:1000, #4060), rabbit anti-mTOR (1:1000, #2983), rabbit anti-p-mTOR (1:1000, #5536), rabbit anti-p-4EBP1 (1:1000, #2855), rabbit anti-4EBP1 (1:1500, #9644), rabbit anti-FOXO3a (1:1000, #12829), rabbit anti-p-FOXO3a (1:1000, #9466) rabbit anti-cleaved PARP (1:1000, #5625) and rabbit anti-cleaved Caspase-3 (1:1000, #9664) were purchased from Cell Signaling Technology; mouse anti-β-actin (1:1000, sc-47778), mouse anti-GAPDH (1:1000, sc-32233) and rabbit anti-PUMA (1:1000, sc-28226) were from Santa Cruz Biotechnology; rabbit anti-p-4EBP1 (1:500, NB100-81769, used in Fig. 1b) and mouse anti-4EBP1 (1:1000, NBP1-47366, used in Fig. 1b) were obtained from Novus Biologicals. The compounds of BH3 mimetics (ABT263 and ABT199) and mTOR inhibitors (BEZ235, AZD8055, and Temsirolimus) were from AbMole Bioscience.

Li H., Liu L., Chang H., Zou Z, & Xing D. (2018). Downregulation of MCL-1 and upregulation of PUMA using mTOR inhibitors enhance antitumor efficacy of BH3 mimetics in triple-negative breast cancer. Cell Death & Disease, 9(2), 137.

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Western Blot Analysis of Apoptosis Markers

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T cells with a >95% purity were purified using an EasySep mouse T cell enrichment kit (StemCell Technologies). T cell lysates were prepared in sample buffer (50 mM Tris-Cl [pH 6.8], 50mM 2-ME, 2% SDS, 0.2% bromophenol blue, and 10% glycerol). Antibodies used for Western blots were rabbit anti-LC3 (polyclonoal P014, MBL International), hamster anti-Bcl-2 (polyclonal, BD pharmingen), rabbit anti-Bcl-x_L (polyclonal), rabbit anti-Mcl-1 (polyclonal, Rockland Immunochemicals), rabbit anti-Bim (polyclonal, Cell Signaling), rabbit anti-Bax (polyclonal, Cell Signaling), rabbit anti-Bak (polyclonal, Cell Signaling), rabbit anti-Bid (polyclonal, Abcam), rabbit anti-PARP-1 (polyclonal, Cell Signaling), rabbit anti-COX IV (polyclonal, Cell Signaling), mouse anti-cytochrome c (7H8.2C12, BioLegend), mouse anti-α-Tubulin (B-5-1-2, Sigma) and goat anti-β-Actin (polyclonal, Santa Cruz Biotechnology). For HRP-labeled western blot, the secondary antibodies were anti-rabbit IgG-HRP, anti-mouse IgG-HRP, anti-hamster IgG-HRP and anti-goat IgG-HRP (Jackson Immunoresearch). The development of the western blot was achieved with SuperSignal West Pico Chemiluminescent substrate (Thermo Scientific). For fluorescent western blot, the secondary antibodies were anti-rabbit IgG-Alexa Fluor 680, anti-mouse IgG-Alexa Fluor 680, and anti-goat IgG-Alexa Fluor 790 (Molecular Probes, Invitrogen).

He M.X, & He Y.W. (2015). c- FLIP protects T lymphocyte from apoptosis in the intrinsic pathway. Journal of immunology (Baltimore, Md. : 1950), 194(7), 3444-3451.

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Mitochondrial Dysfunction and Apoptosis in Retina

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The retinas were collected at 1, 3, 5, and 7 days after the intravitreal injection of mtDNA or control solution. The retinas processed with the Mitochondria Isolation Kit for Tissue as previously described [4 (link)] and the proteins collected from the supernatant and sediment were used for the detection of cytochrome c. Western blotting was performed according to methods described in previous study [4 (link)]. The following primary antibodies were used: rabbit anti-BAX (#2772, Cell Signaling Technology), rabbit anti-BAK (#12105, Cell Signaling Technology), rabbit anti-caspase 9 (AF6348, Affinity Biosciences Ltd), rabbit anti-cleaved caspase 9 (AF5240, affbiotech), rabbit anti-caspase 3 (ab44976, Abcam), rabbit anti-cleaved caspase 3 (ab49822, Abcam), anti-β-actin (ab8227, Abcam), anti-cGAS (ab179785, Abcam), rabbit anti-STING (D1V5L) (#50,494, Cell Signaling Technology), rabbit anti-TBK1/NAK (D1B4) (#3504, Cell Signaling Technology), rabbit anti-phospho-TBK1/NAK (Ser172) (D52C2) XP^® (#5483, Cell Signaling Technology), rabbit anti-IRF3 (D83B9) (#4302, Cell Signaling Technology), rabbit anti-phospho-IRF3 (Ser396) (D6O1M) (#29047, Cell Signaling Technology), anti-interferon β (ab140211, Abcam), rabbit anti-cytochrome c (10993-1-AP, Proteintech), and rabbit anti-VDAC1 (ab154856; Abcam). Three eyes from each group were tested.

Guo Y., Gan D., Hu F., Cheng Y., Yu J., Lei B., Shu Q., Gu R, & Xu G. (2022). Intravitreal injection of mitochondrial DNA induces cell damage and retinal dysfunction in rats. Biological Research, 55, 22.

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Evaluating Apoptosis Markers in Leukemic Cells

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HL60 and HL-60R cells were seeded at a density of 1.5 × 10⁵ cells/well in six-well plates for 36 h. Next, the cells were treated with or without 10 μM MK-4, MKH-DMG, or MKH-SUC for 24, 48, or 72 h. The cells were washed with PBS after removing the drug-containing medium and lysed with RIPA buffer (0.5% NP-40, 0.25% sodium deoxycholate, 0.05% SDS, 150 mM NaCl, and 50 mM HEPES, pH 7.4) containing a protease inhibitor cocktail (Nacalai Tesque, Kyoto, Japan). Each cell lysate was combined with sample loading buffer (Nacalai Tesque), separated on 15% SDS-PAGE gels (SuperSep Ace, FUJIFILM Wako), and transferred onto PVDF membranes (BIO-RAD, Hercules, CA, USA). After blocking, the membranes were incubated with the following primary antibodies: anti-pro/p17-caspase-3, anti-cleaved PARP1 (1:2000) (ab136812; Abcam, Cambridge, UK), rabbit anti-Bak (1:2000) (#12105; Cell Signaling Technology, Danvers, MA, USA), and mouse anti-GAPDH antibody (1:2000) (G8795; Sigma-Aldrich, St. Louis, MO, USA). After washing, the membranes were treated with appropriate secondary antibodies and visualized using Immunostar LD or Immunostar Zeta (FUJIFILM Wako).

Yamakawa H., Setoguchi S., Goto S., Watase D., Terada K., Nagata-Akaho N., Toki E., Koga M., Matsunaga K., Karube Y, & Takata J. (2021). Growth Inhibitory Effects of Ester Derivatives of Menahydroquinone-4, the Reduced Form of Vitamin K2(20), on All-Trans Retinoic Acid-Resistant HL60 Cell Line. Pharmaceutics, 13(5), 758.

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Mitochondrial Dysfunction in Retinal Injury

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The retinas were collected at 1, 3, 5, and 7 days after the intravitreal injection of mtDNA or control solution. The retinas processed with the Mitochondria Isolation Kit for Tissue (C3606, Beyotime, Shanghai, China), as previously described [14] and the proteins collected from the supernatant and sediment were used for the detection of cytochrome c. Western blotting was performed according to methods described in previous study [14] . The following primary antibodies were used: rabbit anti-BAX (#2772, Cell Signaling Technology), rabbit anti-BAK (#12105, Cell Signaling Technology), rabbit anticaspase 9 (AF6348, A nity Biosciences Ltd), rabbit anti-cleaved caspase 9 (AF5240, affbiotech), rabbit anti-caspase 3 (ab44976, Abcam), rabbit anti-cleaved caspase 3 (ab49822, Abcam), anti-β-actin (ab8227, Abcam), anti-cGAS (ab179785, Abcam), rabbit anti-STING (D1V5L) (#50494, Cell Signaling Technology), rabbit anti-TBK1/NAK (D1B4) (#3504, Cell Signaling Technology), rabbit anti-phospho-TBK1/NAK (Ser172) (D52C2) XP® (#5483, Cell Signaling Technology), rabbit anti-IRF3 (D83B9) (#4302, Cell Signaling Technology), rabbit anti-phospho-IRF3 (Ser396) (D6O1M) (#29047, Cell Signaling Technology), anti-interferon β (ab140211, Abcam), rabbit anti-cytochrome c (10993-1-AP, Proteintech), rabbit anti-VDAC1 (ab154856; Abcam). Three eyes from each group were tested.

Guo Y., Gan D., Hu F., Cheng Y., Yu J., Lei B., Shu Q., Gu R., & Xu G. (2021). Intravitreal Injection of Mitochondrial DNA Induces Cell Damage and Retinal Dysfunction in Rats.

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