Ab155948

Manufactured by Abcam

Sourced in United States

Ab155948 is a primary antibody designed for the detection of a target protein. The product is suitable for use in immunoassays such as Western blotting, ELISA, and immunohistochemistry. It is supplied in a liquid format.

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Lab products found in correlation

Flag 80010 1 rr, by Proteintech (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Ab194982, by Abcam (1 mentions) Ab8069, by Abcam (1 mentions) Ab18203, by Abcam (1 mentions) Hrp conjugated secondary antibody, by Sangon (1 mentions) Ab210498, by Abcam (1 mentions) Ab205718, by Abcam (1 mentions) Nitrocellulose membrane, by Merck Group (1 mentions) Ab109012, by Abcam (1 mentions) Ecl detection system, by GE Healthcare (1 mentions) Ab9485, by Abcam (1 mentions)

4 protocols using ab155948

Establishment and Characterization of HIV-1 Infection Models

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Human MT4 T cells and Jurkat cells were grown in Roswell Park Memorial Institute (RPMI) 1640 (Gibco, Logan, UT, USA) at 37°C in 5% CO₂. HOS.CD4.CCR5 cells (hereinafter referred to as HOS cells) and 293T cells were grown in complete Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Logan, UT, USA) supplemented with 10% heat-inactivated fetal calf serum at 37°C in 5% CO₂. The cells were subsequently transfected using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions or inoculated with HIV-1 as described previously [25 (link)]. All cells were purchased from the Cell Bank of the Chinese Academy of Science (Shanghai, China).
Antibodies against HIV-1 p24 (sc-69728) and phosphor-p50 (p-p50; sc-271908) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against p50 (14220-1-AP), Actin (66009-1-Ig), LaminB (12987-1-AP), and Flag (80010-1-RR) were purchased from Proteintech (Rosemont, IL, USA). Antibody against TGIF2 (ab155948) was purchased from Abcam (Cambridge, MA, USA). Antibody against Vpr (MA-001-2) was purchased from Cosmo Bio Co., Ltd. (Tokyo, Japan).

Qiao J., Peng Q., Qian F., You Q., Feng L., Hu S., Liu W., Huang L., Shu X, & Sun B. (2021). HIV-1 Vpr protein upregulates microRNA-210-5p expression to induce G2 arrest by targeting TGIF2. PLoS ONE, 16(12), e0261971.

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Western Blotting of Cell Markers

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Western blot was carried out as previously reported.27 The primary antibodies against TGIF2 (ab155948), vimentin (ab8069), N‐cadherin (ab18203) and E‐cadherin (ab194982) were purchased from Abcam (Cambridge, MA, USA) and HRP‐conjugated secondary antibody was obtained from Sangon (Shanghai, China).

Pang L., Cheng Y., Zou S, & Song J. (2019). Long noncoding RNA SNHG7 contributes to cell proliferation, migration, invasion and epithelial to mesenchymal transition in non‐small cell lung cancer by regulating miR‐449a/TGIF2 axis. Thoracic Cancer, 11(2), 264-276.

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ChIP-qPCR Validation of TGIF2 Binding

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For ChIP, 4L cells were used for each immunoprecipitation (Di Micco et al., 2014 (link)). Five micrograms of TGIF2 antibody (Abcam, ab155948) were used per immunoprecipitation. Next, ChIP qPCR was performed to validate pull down sample. Primers designed to expand regions of the genome not anticipated to be pulled down in the ChIP (GAPDH, HPRT1 or FUT8 exon 5/8) served as negative controls. See Supplementary Table S3 for ChIP-qRT-PCR primer sequences.

Agrawal P., Fontanals-Cirera B., Sokolova E., Jacob S., Vaiana C.A., Argibay D., Davalos V., McDermott M., Nayak S., Darvishian F., Castillo M., Ueberheide B., Osman I., Fenyö D., Mahal L.K, & Hernando E. (2017). A systems biology approach identifies FUT8 as a driver of melanoma metastasis. Cancer cell, 31(6), 804-819.e7.

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Protein Extraction and Western Blot Analysis

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For total protein preparation, tissues and cells were treated with precold RIPA buffer (Beyotime) including protease inhibitor (cocktail, Roche, Basel, Switzerland). For immunoblotting, 50 μg protein samples were processed, separated using an SDS‐PAGE system, transferred onto nitrocellulose membranes (Millipore), blocked in 5% skim milk for 2 h, hybridized with primary antibodies against TGIF2 (1:1000, ab155948, Abcam), Beclin1 (1:1000, ab210498, Abcam), p62 (1:1000, ab109012, Abcam), and GAPDH (1:1000, ab9485, Abcam), and incubated with a secondary antibody (ab205718, 1:10000, Abcam). Finally, the ECL detection system (GE Healthcare) was used for visualization, and Image J software (National Institutes of Health, Bethesda, MD, USA) was employed for analysis. Western blot assay was carried out at least three times.

Tao W., Cao C., Ren G, & Zhou D. (2021). Circular RNA circCPA4 promotes tumorigenesis by regulating miR‐214‐3p/TGIF2 in lung cancer. Thoracic Cancer, 12(24), 3356-3369.

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