Ab33906

Manufactured by Abcam

Sourced in United Kingdom

Ab33906 is a lab equipment product from Abcam. It is a core function product, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

Automatically generated - may contain errors

Lab products found in correlation

Ab182858, by Abcam (2 mentions) P53 polyclonal antibody fl393, by Santa Cruz Biotechnology (1 mentions) Ha antibody sc7392, by Santa Cruz Biotechnology (1 mentions) Antibody against flag m2, by Merck Group (1 mentions) Pcdna3.1 circrna mini vector, by Addgene (1 mentions) Bcl2 associated x (bax), by Thermo Fisher Scientific (1 mentions) Bcl 2, by Thermo Fisher Scientific (1 mentions) Snai1, by Thermo Fisher Scientific (1 mentions) Ab6721, by Abcam (1 mentions) N cadherin, by Abcam (1 mentions) Ab8245, by Abcam (1 mentions) Vimentin, by GeneTex (1 mentions) E cadherin, by GeneTex (1 mentions) Ab115529, by Abcam (1 mentions) Ab131442, by Abcam (1 mentions) Ab109520, by Abcam (1 mentions) Total cell protein extraction kit, by Keygen Biotech (1 mentions) Ab2302, by Abcam (1 mentions) Chemiluminescence kit, by Beyotime (1 mentions) Ab76252, by Abcam (1 mentions)

6 protocols using ab33906

Plasmid Constructs for p53 Regulation

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Plasmid of pGL4-p53 was purchased from Promega. The Plasmids of pCDNA3.1-CDR1as was a gift of Dr. Nikolaus Rajewsky. Plasmids of Myc-p53, Myc-MDM2, PCDH-p53 and HA-Ub were a gift of Dr. Zhang Lingqiang. The pCDNA3.1(+) circRNA mini vector was purchased from Addgene (60648). The plasmid of HA-MDM2 was purchased from Sino Biological (HG11206-CY). All expression constructs were verified by DNA Sequencing.
Antibodies against p53 (DO-1) (ab1101), MDM2 (ab3110), or PUMA (ab33906) were purchased from Abcam Inc. The p53 polyclonal antibody (FL393) and the Myc antibody (9E10), and the HA antibody (sc7392) were purchased from Santa Cruz Biotechnology. The antibody against Flag (M2) was purchased from Sigma. The antibody against GAPDH (AC027) and p21 (A11877) were purchased from Abclonal. The antibody against Phospho-Histone H2A.X (Ser139) (2577 s) was purchased from CST.

Lou J., Hao Y., Lin K., Lyu Y., Chen M., Wang H., Zou D., Jiang X., Wang R., Jin D., Lam E.W., Shao S., Liu Q., Yan J., Wang X., Chen P., Zhang B, & Jin B. (2020). Circular RNA CDR1as disrupts the p53/MDM2 complex to inhibit Gliomagenesis. Molecular Cancer, 19, 138.

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Western Blot Analysis of Epithelial-Mesenchymal Transition Markers

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Total protein was extracted from the cells. Approximately 25 μg sample was processed by 12% polyacrylamide gel electrophoresis and then transferred to polyvinylidene fluoride membranes. The membranes were placed in a blocking solution and slowly shaken for 2 h at 37°C. After blocking, the membranes were mixed with primary antibodies to PUMA (ab33906, 1:1000, Abcam, Cambridge, UK), Bax (#33-6400, 1:500, Invitrogen Inc., Carlsbad, CA, USA), Bcl-2 (#MA5-11757, 1:1000, Invitrogen), E-cadherin (GTX100443, 1:500, GeneTex, Inc., Alton Pkwy Irvine, CA, USA), Vimentin (GTX100619, 1:1000, GeneTex), Snai1 (#14-9859-82, 1:500, Invitrogen), N-cadherin (ab70611, 1:2000, Abcam), CDC73 (#PA5-17159, 1:1000, Invitrogen) and GAPDH (ab8245, 1:2000, Abcam) overnight at 4°C. The membranes were then incubated with the secondary antibody to IgG labelled with horseradish peroxidase (ab6721, 1:1000, Abcam) for 2 h at 37°C. Protein signals were visualized using enhanced chemiluminescence. The protein levels were analyzed using ImageJ software. The relative expression of the target protein was obtained by calculating the ratio of optical density (OD) value of the target protein to that of an internal reference.

Wang J., Luo J., Wu X, & Gao Z. (2021). Circular RNA_0000629 Suppresses Bladder Cancer Progression Mediating MicroRNA-1290/CDC73. Cancer Management and Research, 13, 2701-2715.

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Western Blot Analysis of Key Proteins

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Western blotting analysis was performed as previously described 12 (link). Briefly, total proteins that were extracted using a Total Cell Protein Extraction Kit (KeyGen Biotechnology, Nanjing, China) were divided into equal amounts and electrophoresed, transferred onto a nitrocellulose membrane, and blocked with 2% bovine serum albumin. Primary antibodies against MTBP (1:1000; ab115529, Abcam, Cambridge, UK), MDM2 (1:2000; ab16895), p53 (1:1000; ab131442), p21 (1:2000; ab109520), PUMA (1:2000; ab33906), active caspase3 (1:1000; ab2302), and c-myc (1:1000; ab56) were used to detect the expression of these proteins. After washing four times with TBST/0.1% Tween-20, the membranes were incubated with the corresponding secondary antibody. Bands were visualized using a chemiluminescence kit (Beyotime Biotechnology, Beijing, China) and quantified using the ImageJ software (National Institutes of Health, Bethesda, MD, USA).

Song Y., Zhang L., Jiang Y., Hu T., Zhang D., Qiao Q., Wang R., Wang M, & Han S. (2019). MTBP regulates cell survival and therapeutic sensitivity in TP53 wildtype glioblastomas. Theranostics, 9(20), 6019-6030.

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Western Blot Analysis of Apoptosis-Related Proteins

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The cells were harvested and the total proteins were extracted. The proteins were then denatured and subjected to SDS-PAGE (30 μg of total protein content per well). The proteins were then transferred onto a polyvinylidene difluoride (PVDF) membrane by the wet transfer method. The membrane was then incubated with MST1 monoclonal antibodies (mAbs) (ab124787, Abcam), Bcl-2 mAbs (12789-1-AP, Proteintech), p73 (ab40658, Abcam), p53 antibodies (#21086-1, SAB), PUMA antibodies (ab33906, Abcam), phospho-YAP1 (Ser127) mAbs (ab76252, Abcam), caspase-3 mAbs (Cat. #9665, Cell Signaling Technology), Bax polyclonal antibodies (50599-2-lg, Proteintech) Company), YAP polyclonal antibodies (Cat. #4912, Cell Signaling Technology), and subsequently, other antibodies (1:1200 or 1:1500) overnight at 4°C; then, the membrane was treated with horseradish peroxidase-conjugated secondary antibodies (ZDR-5307, Beijing ZSGB-Bio Origene Co., Ltd, China 1:2000) for 1.5 h. Finally, ECL immunodetection was performed in the dark. The gray values of the protein bands were scanned and analyzed using Image-Pro Plus 6.0 gel image analysis software for semi-quantitative analysis using β-actin (internal reference, AF7018, Santa Cruz, CA) as the control.

Li H., Ruan W.J., Liu L.Q., Wan H.F., Yang X.H., Zhu W.F., Yu L.H., Zhang X.L, & Wan F.S. (2019). Impact of Taurine on the proliferation and apoptosis of human cervical carcinoma cells and its mechanism. Chinese Medical Journal, 132(8), 948-956.

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Western Blot Analysis of Cell Lysates

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The cells from each group were collected, and lysates were generated using the KeyGEN kit (KGP2100, Nanjing, China). The protein concentration was assessed by the BCA method (Thermo Fisher Scientific, Waltham, USA). Next, equal amounts of proteins were analyzed by WB using the following antibodies: anti-MDM2 (556353, BD Pharmingen), anti-p53 (DO-1, Santa Cruz), anti-IDO1 (ab55305, Abcam), anti-p21 (ab109199, Abcam), anti-BCL2 (ab182858, Abcam), anti-BCL-XL (ab32370, Abcam), anti-PUMA (ab33906, Abcam), anti-BAX (ab32503, Abcam) and anti-β-actin (P60709, Cell Signaling Technology). The signals were detected using an ECL chemiluminescence detection system (Bio-Rad, USA). The unprocessed WB images were shown in Supplementary Fig. S4.

Sun C., Li M., Zhang L., Sun F., Chen H., Xu Y., Lan Y., Zhang L., Lu S., Zhu J., Huang J., Wang J., Hu Y., Feng Y, & Zhang Y. (2022). IDO1 plays a tumor-promoting role via MDM2-mediated suppression of the p53 pathway in diffuse large B-cell lymphoma. Cell Death & Disease, 13(6), 572.

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Immunohistochemistry for Apoptosis Regulators

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Immunohistochemistry (IHC) was carried out as followed. In brief, rst, the para n-embeded samples were sectioned into 4µm-thick sections and dewaxed in xylene, rehydrated in rinsed graded ethanol solutions. Endogenous peroxidase activity was then blocked with 3% hydrogen peroxide solution for 10 minutes, rinsed in phosphate buffered saline (PBS) 3 times for 5 min each. Then the tissue sections were heated at 100℃ for 5 minutes in citrate (10mmol/L, pH 6.0) solution to retrieve the antigens. After cooling to room temperature, serum blocker was added to block non-speci c antigen, and were incubated with the primary antibody Bcl-2 (ab182858; Abcam, Cambridge, MA, USA, 1:1000 dilution), Bcl-xL (cat#2764, Cell Signaling Technology, USA, 1:3000 dilution), Mcl-1(ab32087; Abcam, Cambridge, MA, USA, 1:200 dilution), Noxa (ab13654; Abcam, Cambridge, MA, USA, 1:2000 dilution), PUMA (ab33906; Abcam, Cambridge, MA, USA, 1:200 dilution) at 4℃ overnight, followed by washing with PBS for three times, biotinylated goat anti-mouse IgG (1:400; Sigma, St. Louis, MO, USA) or goat anti-rabbit IgG (1:400; Sigma) for 30 min at room temperature. Finally, the signal was developed for visualization with 3,3'-diaminobenzidine tetrahydrochloride (DAB, DAKO, GLOSTRUP, Denmark), and all of the slides were counterstained with hematoxylin.

Guo Y., Zhang L., Zhang N., Chen L., Luo Q., Liu M., Yang D, & Chen J. (2022). Bcl-2 and Noxa are potential prognostic indicators for patients with gastroenteropancreatic neuroendocrine neoplasms. Endocrine, 78(1).

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