Ab86809

Manufactured by Abcam

Sourced in United States

Ab86809 is a lab equipment product offered by Abcam. It serves as a tool for researchers in their scientific investigations, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

Automatically generated - may contain errors

Lab products found in correlation

Pvdf membrane, by Merck Group (2 mentions) Ab137827, by Abcam (2 mentions) Ab16051, by Abcam (2 mentions) Ab226977, by Abcam (2 mentions) Ab230883, by Abcam (2 mentions) Ab8245, by Abcam (2 mentions) Ab32072, by Abcam (2 mentions) Bond max stainer, by Leica (1 mentions) P16 clone jc8, by Santa Cruz Biotechnology (1 mentions) Ab134953, by Abcam (1 mentions) Ab195377, by Abcam (1 mentions) Ab194828, by Abcam (1 mentions) Sc 376104, by Santa Cruz Biotechnology (1 mentions) Irdye 800cw goat anti rabbit igg, by LI COR (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Protease and phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) Ab16663, by Abcam (1 mentions) Ab8227, by Abcam (1 mentions)

6 protocols using ab86809

Immunohistochemical Analysis of SOX4 and p16 Expression

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Specimens were fixed in 10% formaldehyde and embedded in paraffin. Three-micrometer-thick sections were cut from the paraffin-embedded tissue blocks and stained with hematoxylin and eosin. Paraffin sections of each tissue sample were used for immunohistochemical staining with antibodies to SOX4 (polyclonal antibody, ab86809, dilution 1:60; Abcam, Cambridge, MA, USA) and p16 (clone: JC8, dilution 1:1000; Santa Cruz Biotechnology, Dallas, TX, USA). Immunohistochemical staining was performed using the automated Bond Max Stainer (Leica Biosystems, Wetzlar, Germany).

Nasu A., Gion Y., Nishimura Y., Nishikori A., Sakamoto M., Egusa Y., Fujita A., Yoshino T, & Sato Y. (2021). Diagnostic Utility of SOX4 Expression in Adult T-Cell Leukemia/Lymphoma. Diagnostics, 11(5), 766.

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Western Blot Analysis of Signaling Proteins

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Cells were lysed using RIPA buffer. The lysate was separated by 10−15% SDS–polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, Billerica, MA). The membrane was blocked with 5% nonfat milk and incubated with anti-LEF-1 (1:1000, ab137827, Abcam), anti-β-catenin (ab16051, 1:1000, Abcam), anti-CCND2(ab230883, 1:1000, Abcam), anti-CyclinD1 (ab226977, 1:1000, Abcam), anti-MYC (ab32072, 1:1500, Abcam), anti-SOX4 (ab86809, 1:1000, Abcam), anti-Ubiquitin (ab134953, 1:1000, Abcam) and anti-GAPDH (ab8245, 1:1000, Abcam) antibodies overnight at 4 °C. Then the membrane was washed with TBST three times and incubated with the secondary antibody (Abcam, Cambridge, MA) for 1.5 h at room temperature. The ECL chromogenic solution was used to display the chemiluminescence of the bands. The Quantity One 4.4.0 software was used for densitometry determination.

Lin S., Zhang W., Shi Z., Tan L., Zhu Y., Li H, & Peng X. (2021). β-Catenin/LEF-1 transcription complex is responsible for the transcriptional activation of LINC01278. Cancer Cell International, 21, 380.

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Immunofluorescence Analysis of Synovial Tissue

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For IF analysis, paraffin-embedded synovial tissues were deparaffinized, rehydrated, permeabilized and subjected to antigen retrieval. The synovial tissues were blocked with 5% bovine serum albumin (BSA) in phosphate-buffered saline (PBS) for 1 h at RT and then incubated with primary antibodies against SMOC2 (sc-376104, Santa Cruz, 1:50, CA), MYO1C (ab194828, Abcam, 1:50), SOX4 (ab86809, Abcam, 1:50), and ALKBH5 (ab195377, Abcam, 1:200) at 4 °C overnight. Nuclei were stained with DAPI, and images were obtained by fluorescence microscopy (Olympus BX53, Japan).
For cell IF analysis, cells growing on glass coverslips were fixed with 3.7% formaldehyde at RT for 15 min and then permeated with 0.1% Triton X-100 in PBS for 10 min. For the detection of pseudopodia formation, FLSs were stained with Alexa Fluor 546 rhodamine-phalloidin (Molecular Probes, Thermo Fisher Scientific, USA), and for the measurement of monomeric G-actin, FLSs were stained with Fluorescent DNase I (Invitrogen, CA). Nuclei were stained with DAPI, and images were obtained by fluorescence microscopy (Olympus BX53, Japan).

Liu D., Li R., Xu S., Shi M., Kuang Y., Wang J., Shen C., Qiu Q., Liang L., Xiao Y, & Xu H. (2022). SMOC2 promotes aggressive behavior of fibroblast-like synoviocytes in rheumatoid arthritis through transcriptional and post-transcriptional regulating MYO1C. Cell Death & Disease, 13(12), 1035.

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ChIP Assay for SOX4 Transcription Factor

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The ChIP assay kit (CST, USA) was used for ChIP assay according to the manufacturer’s instructions. In brief, RA FLSs transfected with siRNA targeting SOX4 (siSOX4-3) or control siRNA were cross-linked with 1% formaldehyde and lysed in the lysis buffer. Then the lysis was sonicated on ice to generate 200 to 500 bp DNA fragments. After centrifugation, the supernatant containing chromatin was immunoprecipitated with anti-SOX4 (ab86809, Abcam) or isotype control IgG antibodies overnight at 4 °C. Immunoprecipitated DNAs were purified and subjected to RT-qPCR analysis. The ChIP primers were listed in Supplementary Table 2.

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Protein Extraction and Western Blot Analysis

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Total proteins were extracted from cultured cells and mouse prostate tissues using radioimmunoprecipitation assay buffer supplemented with protease and phosphatase inhibitors (Thermo Fisher Scientific, Waltham, MA, USA). In total, 15 μg of protein was separated in a 4–20% gradient SDS-polyacrylamide gel and transferred onto nitrocellulose membranes (Bio-Rad Laboratories, Hercules, CA, USA). Nonspecific binding was blocked with 5% nonfat milk in Tris-buffered saline with Tween 20 as recommended for each antibody. The membranes were further incubated with rabbit anti-Sox4 (ab86809; Abcam, Cambridge, MA, USA), anti-Aldoa (ab232786; Abcam), anti-Ccnd1 (ab16663; Abcam), mouse anti-E-cadherin (Cat. no. 202183; ZEN BIO, Chengdu, China), and rabbit anti-CCND1 (Cat. no. 382442; ZEN BIO) antibodies overnight at 4 °C. IRDye 800CW goat anti-rabbit IgG (Cat. no. 926–32211; LI-COR, Lincoln, NE, USA), goat anti-IgG (H + L chain) (mouse) pAb-HRP (Cat. No. 330, MBL, Beijing, China), and goat anti-IgG (H + L chain) (rabbit) pAb-HRP (Cat. No. 458, MBL) were used as the secondary antibodies, while rabbit anti-beta-actin (ab8227; Abcam) and mouse anti-β-tubulin (Cat. no. 200608; ZEN BIO) were used as internal controls.

Ai J., Li J., Su Q., Ma H., He R., Wei Q., Li H, & Gao G. (2021). rAAV-based and intraprostatically delivered miR-34a therapeutics for efficient inhibition of prostate cancer progression. Gene therapy, 29(7-8), 418-424.

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Western Blot Analysis of Wnt Pathway Proteins

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Cells were lysed using RIPA buffer. The lysate was separated by 10 - 15% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene di uoride (PVDF) membrane (Millipore, Billerica, MA). The membrane was blocked with 5% nonfat milk and incubated with anti-LEF-1 (1:1000, ab137827, Abcam), anti-β-catenin (ab16051, 1:1000, Abcam), anti-CCND2(ab230883, 1:1000, Abcam), anti-CyclinD1 (ab226977, 1:1000, Abcam), anti-MYC (ab32072, 1:1500, Abcam), anti-SOX4 (ab86809, 1:1000, Abcam), and anti-GAPDH (ab8245, 1:1000, Abcam) antibodies overnight at 4 • C. Then the membrane was washed with TBST three times and incubated with the secondary antibody (Abcam, Cambridge, MA) for 1.5 h at room temperature. The ECL chromogenic solution was used to display the chemiluminescence of the bands. The Quantity One 4.4.0 software was used for densitometry determination.

Lin S., Zhang W., Shi Z., Tan L., Zhu Y., Li H., & Peng X. (2020). β-catenin/LEF-1 Transcription Complex is Responsible for the Transcriptional Activation of LINC01278.

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