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Ab28842

Manufactured by Abcam
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Ab28842 is a primary antibody designed for the detection of a specific target protein. This product is intended for use in various laboratory applications, such as immunoassays and immunohistochemistry. The core function of this antibody is to specifically bind and detect the target protein, enabling researchers to study its expression and localization within biological samples.

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Lab products found in correlation

P gsk3β ser9, by Cell Signaling Technology (1 mentions) Vinculin, by Cell Signaling Technology (1 mentions) Gsk3β, by Cell Signaling Technology (1 mentions) Pakt s473, by Cell Signaling Technology (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) β actin, by BD (1 mentions) Complete protease inhibitor mixture tablet, by Roche (1 mentions) Enhanced chemiluminescence, by Thermo Fisher Scientific (1 mentions) Pvdf membrane, by Thermo Fisher Scientific (1 mentions) Ab11174, by Abcam (1 mentions) Nxa931, by GE Healthcare (1 mentions) Na934, by GE Healthcare (1 mentions) H00000191 m07a, by Abnova (1 mentions) Sc 2033, by Santa Cruz Biotechnology (1 mentions) Sc 53993, by Santa Cruz Biotechnology (1 mentions) Ab78318, by Abcam (1 mentions) Ab128874, by Abcam (1 mentions) Protease and phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Abcam (1 mentions)

5 protocols using ab28842

1

Comprehensive Western Blot Profiling

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Cep55 (1:1000, In-house raised in rabbit against murine Cep55 (amino acids 55–250) and The Cell Signaling Technology, CEP55 (D1L4H) Rabbit mAb #81693 1:1000), Vinculin (1:2000; 13901 Cell Signaling Technology), β-Actin (1:2000, 612656 BD Pharmingen), β-Catenin (1:1000, 9582 Cell Signaling Technology), GSK 3β (1:1000, 9369 Cell Signaling Technology), pGSK 3β(Ser9) (1:1000, 9322 Cell Signaling Technology), Cleaved Caspase-3 (1:500, 9664 Cell Signaling Technology), pAKT(s473) (1:1000, 4060 Cell Signaling Technology), AKT (1:1000, 9271 Cell Signaling Technology), p Myc (T58) (1:1000, ab28842 Abcam), Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (1:1000, 8814 Cell Signaling Technology), MYC (Y69) (1:1000, Ab32072Abcam).
Rashidieh B., Shohayeb B., Bain A.L., Fortuna P.R., Sinha D., Burgess A., Mills R., Adams R.C., Lopez J.A., Blumbergs P., Finnie J., Kalimutho M., Piper M., Hudson J.E., Ng D.C, & Khanna K.K. (2021). Cep55 regulation of PI3K/Akt signaling is required for neocortical development and ciliogenesis. PLoS Genetics, 17(10), e1009334.
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2

Neuroblastoma Cell Lysate Analysis

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Neuroblastoma cells were lysed in radioimmune precipitation assay buffer containing 50 mm Tris-HCl (pH 7.4), 150 mm NaCl, 1 mm EDTA, 0.1% sodium deoxycholate, 1% Triton X-100, 0.1% sodium dodecyl sulfate, and a mixture of protease inhibitors (Complete, protease inhibitor mixture tablets; Roche Diagnostics) at 4 °C for 30 min. Cell lysates were mixed with SDS-PAGE loading buffer, loaded onto 8–16% gradient Precise Tris-glycine precast gels (Fisher Scientific) and transferred to PVDF membranes (Fisher Scientific). The antibodies used were MYCN (sc-53993, Santa-Cruz Biotechnology, 1:500 dilution), gamma-H2A.X (phospho S139) (ab11174, Abcam, 1:500 dilution), Actin (sc-1616 Santa Cruz Biotechnology, 1:500 dilution), BLM (A300–110A, Bethyl Laboratories, Cambridge Bioscience, 1:1,000 dilution), PKMYT1 (4282S, Cell Signaling, 1:500 dilution), CKS1B (36-6800, Invitrogen 1:500 dilution), SAHH (H00000191-M07A, Abnova, 1:500 dilution), and c-MYC (phospho T58) (ab28842, Abcam, UK, 1:200 dilution). The membranes were then incubated with appropriate HRP-conjugated secondary antibodies: anti-mouse IgG (NXA931, GE Healthcare, Fisher Scientific), anti-rabbit IgG (NA934, GE Healthcare, Fisher Scientific), or anti-goat IgG (sc-2033, Santa Cruz Biotechnology), all at 1:10,000 dilution. Antibody binding was detected by enhanced chemiluminescence (Fisher Scientific).
Chayka O., D'Acunto C.W., Middleton O., Arab M, & Sala A. (2014). Identification and Pharmacological Inactivation of the MYCN Gene Network as a Therapeutic Strategy for Neuroblastic Tumor Cells. The Journal of Biological Chemistry, 290(4), 2198-2212.
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3

Western Blot Analysis of c-Myc and BRD4

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Cells were seeded at 1,000,000 cells/well/2 mL into 6-well plates. After an overnight incubation, cells were treated with GS-626510 for indicated time points for Western blot analysis. Cells were washed with ice-cold PBS post treatment and resuspended in 200 μL with RIPA buffer containing Protease and Phosphatase Inhibitor (ThermoFisher Scientific, cat#78430). Proteins (10 μg) were resolved using SDS-polyacrylamide gels and blotted onto nitrocellulose membranes. Membranes were washed with TBS (140 mmol/L NaCl, 50 mmol/L Tris-HCl; pH 7.2) containing 0.1%Tween20 (TBST) and 5% skimmed milk to block nonspecific protein binding. Membranes were incubated with antibody against c-Myc (1:1000, 5605S, Cell Signaling Technology), BRD4 (1:500, ab128874, Abcam), c-Myc-phsphoT58 (1:500, ab28842, Abcam), c-Myc-phospho S62 (clone 33A12E10) (1:500, ab78318, Abcam), or GAPDH (1:10000) in TBST for overnight at 4°C, washed three times with TBST, and then incubated with the fluorescently-labeled secondary antibody (1:10000) for 1 hour at room temperature. Immunoreactive proteins were scanned and quantified using Odyssey fluorescence imaging system.
Bonazzoli E., Predolini F., Cocco E., Bellone S., Altwerger G., Menderes G., Zammataro L., Bianchi A., Pettinella F., Riccio F., Han C., Yadav G., Lopez S., Manzano A., Manara P., Buza N., Hui P., Wong S., Litkouhi B., Ratner E., Silasi D.A., Huang G.S., Azodi M., Schwartz P.E., Schlessinger J, & Santin A.D. (2018). Inhibition of BET bromodomain proteins with GS-5829 and GS-626510 in Uterine Serous Carcinoma, a biologically aggressive variant of Endometrial Cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(19), 4845-4853.
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4

Protein Expression Analysis by Western Blot

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Cells were lysed in RIPA buffer (Tris pH 7.5, 20 × 10−3m; NaCl, 150 × 10−3m; 1% Nonidet P‐40; 0.5% sodium deoxycholate; EDTA, 1 × 10−3m; 0.1% SDS) containing protease inhibitor cocktail (K272, Biovision, Milpitas, CA) and phosphatase inhibitor cocktail (K282, Biovision). Samples were then resolved by SDS‐PAGE and immunoblotted with the indicated antibodies; CXCR4, 1:500 (ab28842, Abcam); CSF1, 1:1000 (PA5‐42558, Invitrogen); actin, 1:3000 (Abc‐2004, Abclon, Seoul, Korea).
Choi Y.W., Kim Y.H., Oh S.Y., Suh K.W., Kim Y., Lee G., Yoon J.E., Park S.S., Lee Y., Park Y.J., Kim H.S., Park S.H., Kim J, & Park T.J. (2021). Senescent Tumor Cells Build a Cytokine Shield in Colorectal Cancer. Advanced Science, 8(4), 2002497.
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5

Immunoblot Analysis of Protein Interactions

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Total proteins were extracted and performed the immunoblot analyses. Primary antibodies used: MYC (1:1000; NOVUS; NB600‐336SS), pT58‐MYC (1:1000; Abcam; ab28842), pS62‐MYC (1:1000; Abcam; ab78318), TAF10 (1:800; NOVUS; NBP1‐80706), TRIP12 (1:1000; NOVUS; NBP2‐94325), PCNA (1:1000; Thermo Fisher Scientific; MA5‐11358), Ki67 (1:500; Santa Cruz; sc‐23900), Cleaved‐Caspase 3 (1:500; Abcam; ab2302), Cleaced‐PARP1 (1:1000; Abcam; ab4830), p53 (1:500; Thermo Fisher Scientific; MA5‐12557), Flag (1:1000; Beyotime; AF5051), HA (1:1000; Beyotime; AF5057), GFP (1:1000; Beyotime; AG281) and GAPDH (1:1000; Thermo Fisher Scientific; PA1‐987). Lysates were incubated overnight at 4°C with the appropriate antibodies and Protein A/G beads (Thermo Fisher Scientific) for Co‐IP, followed by immunoblots.
Xiong Y., Wang L., Xu S., Fu B., Che Y., Zaky M.Y., Tian R., Yao R., Guo D., Sha Z., Lin F., Lin X, & Wu H. (2023). Small molecule Z363 co‐regulates TAF10 and MYC via the E3 ligase TRIP12 to suppress tumour growth. Clinical and Translational Medicine, 13(1), e1153.
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