Ab140601

Manufactured by Abcam

Sourced in United States, United Kingdom, China

Ab140601 is a monoclonal antibody that can be used for the detection of a specific target protein in biological samples. It is designed for use in various laboratory techniques, such as Western blotting, ELISA, and immunohistochemistry. The core function of this product is to provide a specific and reliable tool for the identification and analysis of the target protein.

Automatically generated - may contain errors

Lab products found in correlation

Ab179434, by Abcam (13 mentions) Ab33915, by Abcam (3 mentions) β actin, by Cell Signaling Technology (3 mentions) Pvdf membrane, by Merck Group (2 mentions) Chloroquine, by MedChemExpress (2 mentions) Ab7780, by Abcam (2 mentions) Ab134953, by Abcam (2 mentions) Ab178681, by Abcam (2 mentions) Bml pw8810, by Enzo Life Sciences (2 mentions) Mg132 s2619, by Selleck Chemicals (2 mentions) Ab134175, by Abcam (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Ab8638, by Abcam (1 mentions) Ab2741, by Abcam (1 mentions) Ab181602, by Abcam (1 mentions) Hnf1α, by Cell Signaling Technology (1 mentions) Hnf4α, by Cell Signaling Technology (1 mentions) P jnk, by Cell Signaling Technology (1 mentions)

38 protocols using ab140601

Detecting Cellular Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies against p38 (#8690), p-P38 (#4511), ERK1/2 (#4695), p-ERK1/2 (#4376), JNK (#9212), p-JNK (#9255), HNF1α (#89670), HNF4α (#3113) were obtained from Cell Signaling Technology (Cell Signaling Technology, Inc., Danvers, MA, USA). Antibodies against OTUD5 (ab254742), HBc (ab8638), HBx (ab2741), GAPDH (ab181602), Ubiquitin linkage-specific K48 (ab140601), Ubiquitin linkage-specific K63 (ab179434) were obtained from Abcam (Abcam, Cambridge, UK).

Lou B., Ma G., Yu X., Lv F., Xu F., Sun C, & Chen Y. (2023). Deubiquitinase OTUD5 promotes hepatitis B virus replication by removing K48-linked ubiquitination of HBV core/precore and upregulates HNF4ɑ expressions by inhibiting the ERK1/2/mitogen-activated protein kinase pathway. Cellular and Molecular Life Sciences, 80(11), 336.

+ Open protocol

+ Expand

Immunoblotting analysis of signaling proteins

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The experiments were performed according to the previous report [9 (link)]. Proteins were quantified using a BCA protein assay kit (Thermo Scientific, Waltham, MA), and 25 μg proteins were separated using 10% SDS-PAGE and transferred to a PVDF membrane (Millipore). The antibodies for USP5 (1 : 500, #ab155993), TRAF6 (1 : 500, #ab33915), p65 (1 : 500, #ab16502), phos-p65 (1 : 500, #ab76302), IκBα (1 : 500, #ab32518), phos-IκBα (1 : 500, #ab133462), β-actin (1 : 1000, #ab179467), Myc (1: 1000, #ab32), Flag (1 : 1000, #ab49763), and K48-ub (1 : 500, #ab140601) were all bought from Abcam (Abcam, Cambridge, USA).

Luo X.B., Xi J.C., Liu Z., Long Y., Li L.T., Luo Z.P, & Liu D.H. (2020). Proinflammatory Effects of Ubiquitin-Specific Protease 5 (USP5) in Rheumatoid Arthritis Fibroblast-Like Synoviocytes. Mediators of Inflammation, 2020, 8295149.

+ Open protocol

+ Expand

Western Blot Analysis of Protein Interactions

Check if the same lab product or an alternative is used in the 5 most similar protocols

As formerly described, the Co-IP, western blotting, GST pull-down as well as in vitro ubiquitination assays were implemented [18 (link), 25 (link)]. For the Western blot analysis, cells or tissues were dissolved in Tris (20 mM, with a pH of 7.8), NaCl (150 mM), 0.1% Triton X-100 with protease inhibitors (phenylmethylsulphonyl fluoride (PMSF) 100 mM, 1 μM pepstatin and 5 μg/ml aprotinin). Subsequently, SDS/PAGE was applied to separate the equal amounts of cell lysates, which were electrotransferred to the membranes of polyvinylidene fluoride (Millipore, USA) and blocked in skim milk (5%). Through using the specific primary antibodies, and then the proper secondary antibodies conjugated with HRP, the membranes were immunoblotted. The immunoreactive bands were observed using a chemiluminescence kit with high sensitivity. The following antibodies were used: antibodies against FAT10 (1:1000, Santa cruz, sc-393630), USP7 (1:1000, Abcam, ab108931), CHK1 (1:1000, CST, #2360), CDKN1A (1:1000, Abcam, ab102013), TGF-β (1:1000, Abcam, ab215715), and CTGF (1:1000, Abcam, ab209780), Flag (1:1000, Sigma, F1804), His (1:1000, Abmart, M30111), ubiquitin (1:1000, Abcam, ab140601) and Tubulin (1:1000, Abcam, ab7291).

Shao Y., Zhang W., Du D., Yu Y., Li Q, & Peng X. (2022). Ubiquitin-like protein FAT10 promotes renal fibrosis by stabilizing USP7 to prolong CHK1-mediated G2/M arrest in renal tubular epithelial cells. Aging (Albany NY), 14(18), 7527-7546.

+ Open protocol

+ Expand

Ubiquitin Immunoprecipitation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

This assay was conducted following previous description (21 (link)). Above all, IP lysis buffer (20-188, Millipore) was utilized for gathering MG132-treated/-untreated M0 cells in advance. To collect the supernatants, the cell lysates were subjected to 20-minute centrifugation at 16,000 × g. Subsequently, the collected supernatants were pre-cleared with protein A/G beads, followed by 2-hour incubation with primary antibody of Ubiquitin (ab140601, Abcam). Then, the immunoprecipitated protein complexes were collected and eluted, followed by transferring to SDS-PAGE gel. Eventually, the eluted samples and input fractions were subjected to western blot analysis.

Guan H., Mao L., Wang J., Wang S., Yang S., Wu H., Sun W., Chen Z, & Chen M. (2022). Exosomal RNF157 mRNA from prostate cancer cells contributes to M2 macrophage polarization through destabilizing HDAC1. Frontiers in Oncology, 12, 1021270.

+ Open protocol

+ Expand

TRIM26 Ubiquitination and GPX4 Regulation in Oxidative Stress

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies and reagents were acquired from designated suppliers:
TRIM26 (ab188017, Abcam), TRIM26 (27013-1-AP, Proteintech), TRIM26 (sc-393832, Santa Cruz Biotechnology), GPX4 (ab125066, Abcam), GPX4 (sc-166570, Santa Cruz Biotechnology), PLK1 (ab189139, Abcam), p-PLK1 (ab155095, Abcam), β-actin (ab8226, Abcam), K48-ubiquitin (ab140601, Abcam), K63-ubiquitin (12930, Cell Signaling Technology), anti-Flag (66008, Proteintech), anti-Myc (16286, Proteintech), p-S/T (61 G, abmart), p-S/T (612549, BD Biosciences), Anti-His (66005, Proteintech), anti-Flag (14793, Cell Signaling Technology), anti-Myc (2276, Cell Signaling Technology), Anti-His (12689, Cell Signaling Technology), anti-HA (3724, Cell Signaling Technology),
MDA (ab27642, Abcam), 4-HNE (ab48506, Abcam), MG132 (S1748, Beyotime), cycloheximide (HY12320, MedChemExpress), Chloroquine (HY-17589A, MedChemExpress), Onvansertib (HY-15828, MedChemExpress), MLN0905 (HY-15155, MedChemExpress), Recombinant Human PLK1 (ab271716, Abcam).

Wang Z., Xia Y., Wang Y., Zhu R., Li H., Liu Y, & Shen N. (2023). The E3 ligase TRIM26 suppresses ferroptosis through catalyzing K63-linked ubiquitination of GPX4 in glioma. Cell Death & Disease, 14(10), 695.

+ Open protocol

+ Expand

Co-immunoprecipitation of p73 and ubiquitin

Check if the same lab product or an alternative is used in the 5 most similar protocols

For Co-IP assay, magnetic beads were pre-incubated with the antibody of P73 (1:100, ab202474, Abcam). After that, the cells were lysed in with Co-IP buffer and centrifuged. The supernatant was collected and incubated with the beads overnight at 4 ℃. Finally, the beads were collected and the protein levels in the immunoprecipitates were examined by WB analysis using anti-UB antibody (1:1000, ab140601, Abcam).

Wang Y., Liu M., Liu X, & Guo X. (2023). LINC00963-FOSB-mediated transcription activation of UBE3C enhances radioresistance of breast cancer cells by inducing ubiquitination-dependent protein degradation of TP73. Journal of Translational Medicine, 21, 321.

+ Open protocol

+ Expand

Detecting K48 and K63 Polyubiquitination

Check if the same lab product or an alternative is used in the 5 most similar protocols

The cell lysates were prepared, and 10 µg of TRAF6 (Santa Cruz Biotechnology # sc‐8409) antibodies and 1 mg of each protein sample were used for co‐IP assays (Thermo Scientific, USA) to detect K48‐specific (1:1000, Abcam # ab140601) and K63‐specific (1:1000, Abcam # ab179434) polyubiquitination. The eluted protein samples analysed by Western blot.

Yan K., Wu C., Ye Y., Li L., Wang X., He W., Ren S, & Xu Y. (2020). A20 inhibits osteoclastogenesis via TRAF6‐dependent autophagy in human periodontal ligament cells under hypoxia. Cell Proliferation, 53(3), e12778.

+ Open protocol

+ Expand

Investigating USP2A Interactions with Transcription Factors

Check if the same lab product or an alternative is used in the 5 most similar protocols

The coding region of human USP2A (accession number NM_004205) without the stop codon was cloned into pcDNA3.2/V5-DEST (Thermo Fisher Scientific) using BP clonase II (Thermo Fisher Scientific). The resultant plasmid encoded C-terminal V5-tagged USP2A. Halo-tagged Oct-1 and Oct-2 expression plasmids were purchased from Promega (Madison, WI). The HA-tagged ubiquitin plasmid was purchased from Addgene (Cambridge, MA). The USP2A, or control pcDNA3.2/V5-DEST plasmid, plasmids encoding Halo-tagged Oct-1 or Oct-2 and HA-tagged ubiquitin were transfected into HEK293FT cells using Lipofectamine 2000 reagent (Thermo Fisher Scientific). The pull-down and elution of Oct proteins were performed using Magne HaloTag beads (Promega) and HaloTEV protease (Promega) as per the manufacturer's instructions. For detection of Western blot bands, corresponding to K48- and K63-linked ubiquitin chains, Oct proteins, HA-tagged ubiquitin, and V5-tagged USP2, 1000-fold-diluted antibodies against polyubiquitin chains formed by K48 residues (ab140601; Abcam) and K63 residues (ab179434; Abcam), Oct-1 (A301-717A; Bethyl Laboratories), Oct-2 (ab179808; Abcam), HA-tag (#3724; CST), and V5-tag (A190-120F; Bethyl Laboratories) were used as the primary antibodies.

Kitamura H., Ishino T., Shimamoto Y., Okabe J., Miyamoto T., Takahashi E, & Miyoshi I. (2017). Ubiquitin-Specific Protease 2 Modulates the Lipopolysaccharide-Elicited Expression of Proinflammatory Cytokines in Macrophage-like HL-60 Cells. Mediators of Inflammation, 2017, 6909415.

+ Open protocol

+ Expand

TRAF6 Immunoprecipitation and Polyubiquitin Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunoprecipitation of TRAF6 was performed using an anti-TRAF6 (ab33915; Abcam) antibody with a Pierce Crosslink IP Kit (Thermo Fisher Scientific) according to the manufacturer's instructions. For detection of Western blot bands, TRAF6 (ab33915; Abcam), polyubiquitin chains formed by K63-linked (ab179434; Abcam) and K48-linked (ab140601; Abcam) primary antibodies were used (1000-fold dilution).

+ Open protocol

+ Expand

Comprehensive Immunoblotting Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunoblotting assays were carried out as previously described⁹ with the following primary Abs: DAB2IP (23582‐1‐AP, Proteintech), c‐Myc (ab32072, Abcam), c‐Myc (67447‐1‐Ig, Proteintech), p‐c‐Myc^S62 (ab185656, Abcam), p‐c‐Myc^T58 (ab185655, Abcam), glycogen synthase kinase 3β (GSK3β; 10044‐T32, SinoBiological), p‐GSK3β^S9 (#5558, CST), Bcl‐2 (12789‐1‐AP, Proteintech), Bax (#5023, CST), B56α (ab89621, Abcam), Ubiquitin (10201‐2‐AP, Proteintech), Ubiquitin (linkage‐specific K48) (ab140601, Abcam), Nanog (#8822S, CST), CD44 (60224‐1‐Ig, Proteintech), CD133 (18470‐1‐AP, Proteintech), and GAPDH (60004‐1‐Ig, Proteintech). Secondary Abs conjugated to HRP (Servicebio) were used, followed by chemiluminescence.

Li H., Zhou Y., Wang M., Wang H., Zhang Y., Peng R., Zhang R., Zhang M., Zhang M., Qiu P., Liu L., Zhao Q, & Liu J. (2021). DOC‐2/DAB2 interactive protein destabilizes c‐Myc to impair the growth and self‐renewal of colon tumor‐repopulating cells. Cancer Science, 112(11), 4593-4603.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!