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Anti e1a

Manufactured by Santa Cruz Biotechnology
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Anti-E1A is a lab equipment product offered by Santa Cruz Biotechnology. It is a reagent used for the detection and study of the E1A protein, which is an early gene product of adenovirus. The core function of Anti-E1A is to provide a specific and reliable tool for researchers investigating the role and regulation of the E1A protein in various biological systems.

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Lab products found in correlation

Anti mouse proliferating cell nuclear antigen pcna, by Agilent Technologies (2 mentions) Anti actin, by Merck Group (2 mentions) Anti hras, by Santa Cruz Biotechnology (2 mentions) Anti cyclin a2, by Merck Group (2 mentions) Anti hmga2, by Santa Cruz Biotechnology (2 mentions) Anti β actin, by Merck Group (2 mentions) 0.45 μm nitrocellulose membrane, by Merck Group (1 mentions) Cell lysis buffer, by Beyotime (1 mentions) Caspase 8, by Cell Signaling Technology (1 mentions) Bcl2 associated x (bax), by Santa Cruz Biotechnology (1 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Anti bax, by Cell Signaling Technology (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Nbt bcip color substrate, by Promega (1 mentions) Anti mcl 1, by Cell Signaling Technology (1 mentions) Anti bcl xl, by Cell Signaling Technology (1 mentions) Anti caspase 3, by Cell Signaling Technology (1 mentions) Anti histone h3, by Abcam (1 mentions)

Spelling variants of this product

Anti–Adenovirus-2/5 E1A polyclonal antibody (2 citations) Rabbit anti-E1A IgG Ab (2 citations) Anti-Ad2/5 E1A antibody (2 citations) Adenovirus rabbit anti-E1A (2 citations)

10 protocols using anti e1a

1

Intratumoral Injection and Analysis

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Example 11

PBS with an MGF, HmT (5×1010 VP) with an MGF, or HmT-PCION (5×1010 VP) with or without an MGF was intratumorally injected and, 72 hours after injection, MCF7 tumor tissues were obtained. The obtained tumor tissues were fixed in 10% formalin and paraffin-embedded, and 5 μm sections were stained with hematoxylin and eosin and analyzed using a microscope. The tumor sections were also stained with an anti-mouse proliferating cell nuclear antigen (PCNA, available from DaKo) which indicates tumor cell proliferation or an anti-E1A (Santa Cruz Biotechnology) antibody which indicates Ad replication in tumors. Immunohistochemical sections were counterstained with Mayer's hematoxylin.

US11000590B2. Virus-PCION complex having enhanced antitumor effect by using electromagnetic field (2021-05-11). INDUSTRY-UNIVERSITY COOPERATION FOUNDATION HANYANG UNIVERSITY [KR]. Inventors: Chae Ok Yun [KR], Joung-Woo Choi [KR].
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2

Protein Expression Analysis by Western Blot

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Total proteins were collected by cell lysis buffer (Beyotime, Nantong, China). Protein concentrations were determined by Pierce BCA protein assay kit (ThermoFisher scientific). Proteins were separated by SDS-PAGE on an 8–12% gel and then transferred to a 0.45 μm nitrocellulose membrane (Millipore Corp., Bedford, MA, USA). After being blocked in blocking buffer (5% bovine serum albumin, 10 mmol/L Tris–HCl, pH 8.0, 150 mmol/L NaCl, and 0.05% Tween20), the membranes were incubated with primary antibodies and the corresponding secondary fluorescent antibodies (LI-COR Biosciences Inc., Lincoln, NE, USA). The fluorescent signal was detected by the Odyssey infrared imaging system (LI-COR Biosciences Inc.). The primary antibody anti-E1A, TRAIL, Bax, and GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). XIAP, PARP, and caspase-8 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).
Wang Y., Wang B., Liang J., Cui C., Ying C., Huang F., Ma B., Zhou X, & Chu L. (2019). Oncolytic viro-chemotherapy exhibits antitumor effect in laryngeal squamous cell carcinoma cells and mouse xenografts. Cancer Management and Research, 11, 3285-3294.
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3

Western Blot Analysis of Apoptosis Markers

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The cells were harvested from the plates, and aliquots of cell extracts were separated on a 12% SDS–polyacrylamide gel. The proteins were then transferred onto a nitrocellulose membrane and incubated overnight at 4 °C with the following rabbit polyclonal antibodies: anti-IL-24 (Gen Hunter C-corporation, Nashville, TN, USA), anti-caspase-3, anti-Bcl-xl, anti-Bax, anti-Mcl-1 (Cell Signaling, Beverly, MA, USA), anti-β-actin, and anti-E1A antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), respectively. Membranes were then washed and incubated with alkaline phosphatase-conjugated secondary antibodies in tris-buffered saline containing Tween-20 for 2 h and developed using the NBT/BCIP color substrate (Promega). The densities of the bands on the membrane were scanned and analysed with ImageJ (LabWorks Software, UVP Upland, CA, USA).
Jiang G., Yang C.S., Xu D., Sun C., Zheng J.N., Lei T.C, & Liu Y.Q. (2014). Potent anti-tumour activity of a novel conditionally replicating adenovirus for melanoma via inhibition of migration and invasion. British Journal of Cancer, 110(10), 2496-2505.
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4

Immunoblotting analysis of key proteins

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The following antibodies were used for immunoblotting: anti-HRAS (Santa Cruz, sc-29), anti-human p21 (Santa Cruz, sc-397), anti-E1A (Santa Cruz, sc-430); anti-ß-actin (Sigma A5441), anti-Cyclin A2 (Sigma C4710), anti-human p53 (DO-1, Sigma P6874), anti-MDM2 (clones 2A10 and 4B11) [18 (link)], anti-Histone H3 (Abcam ab1791), anti-HMGA2 (Santa Cruz, sc-30223), anti-SCD/Scd1 (Cell Signaling, #2438), anti-mouse p53 (Biovision #3036) and anti-mouse p21 (Santa Cruz #sc-6246), anti-α-Tubulin (Abcam #Ab18251), anti-SREBP1 (Santa Cruz, sc-13551). Immunoblotting analysis was carried out as described [18 (link)].
Kirschner K., Samarajiwa S.A., Cairns J.M., Menon S., Pérez-Mancera P.A., Tomimatsu K., Bermejo-Rodriguez C., Ito Y., Chandra T., Narita M., Lyons S.K., Lynch A.G., Kimura H., Ohbayashi T., Tavaré S, & Narita M. (2015). Phenotype Specific Analyses Reveal Distinct Regulatory Mechanism for Chronically Activated p53. PLoS Genetics, 11(3), e1005053.
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5

ErbB2/ErbB3 Expression Analysis in Breast Cancer

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At 48 h post treatment with replication-deficient Ad (dAd) or oncolytic Ad (oAd or oAd/shErbB3), BT474 cells were harvested and lysed in RIPA buffer (Elpis biotech, Seoul, Korea). The precleared lysates were separated by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride membrane. The membranes were blocked at room temperature for 1 h, followed by incubation at room temperature for another 1 h with the following antibodies: anti-ErbB2, anti-ErbB3, or anti-E1A. All antibodies, except anti-E1A, were purchased from Cell Signaling Technology (Beverly, MA, USA), and anti-E1A was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All membranes were subsequently incubated at room temperature for 1 h with horseradish peroxidase (HRP)-conjugated secondary antibodies, goat anti-rabbit IgG HRP or goat anti-mouse IgG HRP (Cell Signaling Technology). Anti-β-actin (Cell Signaling Technology) was used as an internal loading control. Finally, the blots were developed using enhanced chemiluminescence (ECL) (Pierce, Rockford, IL, USA), imaged using the LAS4000 Luminescent Image Analyzer (Fujifilm Corp., Tokyo, Japan).
Jung B.K., Kim Y.J., Hong J., Chang H.G., Yoon A.R, & Yun C.O. (2022). ErbB3-Targeting Oncolytic Adenovirus Causes Potent Tumor Suppression by Induction of Apoptosis in Cancer Cells. International Journal of Molecular Sciences, 23(13), 7127.
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6

Protein Expression Analysis of Adenoviral Infection

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Cells were seeded in 6-well plates and infected with OAd. DD3.mK5 at 5 MOI. After 48h, the cells were harvested. The protein concentration levels were measured by the Lowry assay (BioRad, Hercules, CA, USA) using manufacturer’s instructions. Western blot was performed according to standard protocols [10 (link)]. Antibodies information as follows: anti-GAPDH (CWBIO, Beijing, China); anti-E1A (Santa Cruz biotechnology, Santa Cruz, CA, USA).
Hao J., Xie W., Li H, & Li R. (2018). Prostate Cancer-Specific of DD3-driven Oncolytic Virus-harboring mK5 Gene. Open Medicine, 14, 1-9.
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7

Antibody Dilutions for Western Blot

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Anti-tubulin (Dilution 1/10,000 for western blot WB) was purchased from SIGMA (Catalog number T5168), Anti-beta actin (Dilution 1/10,000 for western blot WB) was purchased from SIGMA (Catalog number A5316), anti-E1A (dilution 1/1000 for WB and 1/200 for immunofluorescence IF) from Santa Cruz Biotechnology (Catalog number sc-58658, lot # G2611), anti-CGBP (Cfp1; dilution 1/1000 for WB) from Santa Cruz Biotechnology (Catalog number sc-136419, lot # H0510), HBc (Dilution 1/30,000 for WB and 1/500 for IF) was from DAKO (Catalog number B0586, lot # 1018412) and prediluted anti-HBs (no dilution) from sigma (Catalog number ab859, lot # GR3199714-3).
Moreau P., Cournac A., Palumbo G.A., Marbouty M., Mortaza S., Thierry A., Cairo S., Lavigne M., Koszul R, & Neuveut C. (2018). Tridimensional infiltration of DNA viruses into the host genome shows preferential contact with active chromatin. Nature Communications, 9, 4268.
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8

Intratumoral Injection and Histological Analysis

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Example 11

PBS with an MGF, HmT (5×1010 VP) with an MGF, or HmT-PCION (5×1010 VP) with or without an MGF was intratumorally injected and, 72 hours after injection, MCF7 tumor tissues were obtained. The obtained tumor tissues were fixed in 10% formalin and paraffin-embedded, and 5 μm sections were stained with hematoxylin and eosin and analyzed using a microscope. The tumor sections were also stained with an anti-mouse proliferating cell nuclear antigen (PCNA, available from DaKo) which indicates tumor cell proliferation or an anti-E1A (Santa Cruz Biotechnology) antibody which indicates Ad replication in tumors. Immunohistochemical sections were counterstained with Mayer's hematoxylin.

US10357564B2. Virus-PCION complex having enhanced antitumor effect by using electromagnetic field (2019-07-23). INDUSTRY-UNIVERSITY COOPERATION FOUNDATION HANYANG UNIVERSITY [KR]. Inventors: Chae Ok Yun [KR], Joung-Woo Choi [KR].
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9

Ectopic Gene Expression and Immunoblotting

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Ectopic genes were introduced to cells by retrovirus-mediated gene transfer (Young et al. 2009 (link)) using the retroviral vectors pLNCX2-Neo (ER:H-RASG12V) and pWZL-Hygro (E1A). Immunoblotting (Young et al. 2009 (link)) utilized the following antibodies: anti-H-RAS, anti-E1A and anti-HMGA2 (Santa Cruz); anti-ß-actin, anti-cyclin A2, and anti-p53 (Sigma); anti-PARP (Cell Signaling).
Madhu B., Narita M., Jauhiainen A., Menon S., Stubbs M., Tavaré S., Narita M, & Griffiths J.R. (2015). Metabolomic changes during cellular transformation monitored by metabolite–metabolite correlation analysis and correlated with gene expression. Metabolomics, 11(6), 1848-1863.
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10

Comprehensive Immunoblotting Assay Protocol

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Immunoblotting was performed as previously described 19 . The following antibodies were used for immunoblotting: anti-beta-Actin (Sigma, A5441, 1:5000), anti-Cyclin A (Sigma, C4170, 1:1000), anti-Involucrin (Sigma, I9018, 1:1000), anti-p16 (H-156, Santa Cruz, sc-759, 1:500) and (G175-1239, BD Pharmingen, 554079, 1:500), anti-p21 (F-5, Santa Cruz, sc-6246, 1:500), anti-p53 (DO-1, Sigma, P6874, 1:1000), anti-C/EBPβ (C-19, Santa Cruz, sc-150, 1:500), anti-c-H-Ras (F235-1.7.1, Calbiochem, OP23, 1:500), anti-E1A (Santa Cruz, sc-430, 1:2000), anti-HMGA1 (Active Motif, 39615, 1:2000). The anti-Pan-LCE2 antibody (1:1000) was raised in rabbits against a synthetic peptide (RPRLFHRRRHQSPD), as previously described (note, this antigen sequence perfectly matches human LCE2B, C and D, and differs by one amino acid for LCE2A) 26 . The following antibodies were used for ChIP: anti-H3K9me3 (clone
Tomimatsu K., Bihary D., Olan I., Parry A.J., Schoenfelder S., Chan A.S.L., Slater G.S.C., Ito Y., Rugg-Gunn P.J., Kirschner K., Bermejo-Rodriguez C., Seko T., Kugoh H., Shiraishi K., Sayama K., Kimura H., Fraser P., Narita M., Samarajiwa S.A, & Narita M. (2022). Locus-specific induction of gene expression from heterochromatin loci during cellular senescence. Nature aging, 2(1).
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