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Ab21981

Manufactured by Abcam
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Sourced in United Kingdom

Ab21981 is a lab equipment product offered by Abcam. It is designed for use in scientific research applications. The core function of this product is to provide a tool for researchers to facilitate their experiments and laboratory work, but a detailed description of its specific capabilities or intended use is not available.

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Lab products found in correlation

Nbp1 54578, by Novus Biologicals (2 mentions) Ab220867, by Abcam (2 mentions) Ab109489, by Abcam (2 mentions) Spss 22.0 statistical software, by IBM (2 mentions) Ripa buffer, by Thermo Fisher Scientific (2 mentions) Ab32136, by Abcam (1 mentions) Dynabeads protein g, by Thermo Fisher Scientific (1 mentions) Ma1 065, by Thermo Fisher Scientific (1 mentions) Leica tcs sp5 aobs laser scanning confocal microscope, by Leica Microsystems (1 mentions) Leica 63 n a 1.4 oil objective, by Leica camera (1 mentions) Ab215206, by Abcam (1 mentions) Ab169755, by Abcam (1 mentions) Ab135669, by Abcam (1 mentions) Ab181602, by Abcam (1 mentions) Ab32364, by Abcam (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Phosphatase inhibitor cocktail, by Roche (1 mentions) Ab112501, by Abcam (1 mentions) Ab4821, by Abcam (1 mentions) Ab31828, by Abcam (1 mentions)

7 protocols using ab21981

1

Clathrin and AP2 Immunoprecipitation Assay

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For the Clathrin and AP2 immunoprecipitation (IP) assays, protein samples were added to Dynabeads-Protein G (30 μg/100 μL) according to the manufacturer protocol (Invitrogen, DK).
The following were used for both WB and IP analysis: rabbit monoclonal (Y188) antibody to APP (ab32136, Abcam, UK), mouse anti-alpha adaptin antibody (AP6) specific against AP2 (MA1-064, Thermo Fisher, DK), mouse anti-Clathrin heavy chain antibody (clone X22) (MA1-065, Thermo Fisher, DK). Rabbit, anti-AP1+2 antibody (ab21981) were provided by Abcam.
APP pTyr residues were immunoprecipitated using anti-pTyr antibody clone 4G10® agarose conjugate (clone 16-10, Millipore) and analyzed with rabbit anti-APP (clone Y188) following the same procedures previously reported (Matrone et al., 2011 (link)). Rabbit anti-pan Fyn (#4023) and anti-Src pTyr416 (#2101) and anti-Src pTyr527 (#2105) were provided by Cell Signaling Technology (BioNordika, DK).
Poulsen E.T., Iannuzzi F., Rasmussen H.F., Maier T.J., Enghild J.J., Jørgensen A.L, & Matrone C. (2017). An Aberrant Phosphorylation of Amyloid Precursor Protein Tyrosine Regulates Its Trafficking and the Binding to the Clathrin Endocytic Complex in Neural Stem Cells of Alzheimer's Disease Patients. Frontiers in Molecular Neuroscience, 10, 59.
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2

Quantifying Pulmonary Inflammation via H&E and IHC

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To estimate the degree of inflammation, the lung tissues fixed with paraformaldehyde (4% v/v) were stained with H&E (Sigma-Aldrich, St. Louis, MO, USA) after going through a series of procedures. The degree of inflammation was quantified using an image analyser (IMT i-Solution, Vancouver, BC, Canada). The IHC examinations were performed as previously described [20 (link)]. The anti-AP-1 (ab21981; 1:200; Abcam) and anti-TXNIP (NBP1-54578; 1:200; Novus Biologicals, Littleton, CO, USA) were used as primary antibodies.
Lim J.O., Lee S.J., Kim W.I., Pak S.W., Kim J.C., Kim J.S., Cho Y.K., Lee I.C, & Shin I.S. (2021). Melatonin Alleviates Silica Nanoparticle-Induced Lung Inflammation via Thioredoxin-Interacting Protein Downregulation. Antioxidants, 10(11), 1765.
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3

Double-Immunofluorescence Imaging of AP-1 and TXNIP

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Double-immunofluorescence was performed as previously described [23 (link)], using anti-AP-1 (ab21981; 1:200; Abcam) and anti-TXNIP (NBP1-54578; 1:200; Novus Biologicals) antibodies, and imaging was completed using a Leica TCS SP5 AOBS laser scanning confocal microscope (Leica Microsystems, Hesse, Germany) under a Leica 63× (N.A. 1.4) oil objective.
Lim J.O., Lee S.J., Kim W.I., Pak S.W., Kim J.C., Kim J.S., Cho Y.K., Lee I.C, & Shin I.S. (2021). Melatonin Alleviates Silica Nanoparticle-Induced Lung Inflammation via Thioredoxin-Interacting Protein Downregulation. Antioxidants, 10(11), 1765.
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4

Protein Isolation and Analysis in Lung Tissue

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Total proteins were isolated from right lung tissue (50 µg) and cells (5×106) using RIPA buffer (#89900; Thermo Scientific, Worcester, MA, USA) containing a protease inhibitor cocktail (#P2714; Sigma) and phosphatase inhibitor cocktail (#04906845001; Roche Applied Science, Indianapolis, IN, USA). Homogenates were centrifuged at 2,834 g at 4 °C for 20 min before the supernatant was collected. The total protein concentrations were determined by bicinchoninic acid assay (#23227, PierceTM Biotechnology, Rockford, IL, USA). Standard SDS-PAGE technique (27 (link)) was performed on equal amounts of proteins, with antibodies of p-STAT5 (ab32364; 1:500), RORγt (ab135669; 1:500), FOXP3 (ab215206; 1:500), JNK (ab112501; 1:500), p-JNK (ab4821; 1:500), arachidonate 5-lipoxygenase (Alox5; ab169755; 1:500), activator protein 1 (AP-1, ab21981; 1:500), and GAPDH (ab181602; 1:500) obtained from Abcam. The band intensity was measured by ImageJ2× 2.1.4.7 (Wayne Rasband, National Institutes of Health, USA).
Wei C., Huang L., Zheng Y, & Cai X. (2021). Selective activation of cannabinoid receptor 2 regulates Treg/Th17 balance to ameliorate neutrophilic asthma in mice. Annals of Translational Medicine, 9(12), 1015.
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5

Western Blot Analysis of Signaling Proteins

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The cells were lysed with radioimmunoprecipitation assay buffer at 4°C, and then separated by high speed centrifugation (14,000 rpm) at 4°C for 10 min. Next, the supernatant was quantified using the BCA Protein Assay Kit (Shanghai Biotech Biotechnology Research Institute). The protein sample was then loaded onto 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis at 70 V for 30 min then 120 V for 90 min and transferred to a nitrocellulose membrane (Millipore, Boston, MA, USA) at 300 mA for 2 h. After blocking with 5% skim milk, the blot was probed with a primary antibody. After washing with phosphate buffered saline, the horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin G secondary antibodies (ab150077, 1:1,000, Abcam, Cambridge, UK) were added. The primary antibodies used in this study include rabbit anti-MAP2K3 antibody (ab195037, 1:500, Abcam), rabbit anti-AP-1 antibody (ab21981, 1:200, Abcam), and rabbit anti-p38 antibody (ab31828, 1:300, Abcam). We then uniformly add the chemiluminescence reagent to the membrane and develop with the developer. The expression level of the protein was analyzed by chemiluminescence and quantified by Image J software (Version 1. 46; National Institutes of Health, Bethesda, MD, USA). We used GAPDH as an endogenous control to normalize protein expression.
Wang J, & Zhao Q. (2020). Linc02381 Exacerbates Rheumatoid Arthritis Through Adsorbing miR-590-5p and Activating the Mitogen-Activated Protein Kinase Signaling Pathway in Rheumatoid arthritis-fibroblast-like synoviocytes. Cell Transplantation, 29, 0963689720938023.
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6

Chromatin Immunoprecipitation and Quantification

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HMCs were cultured to 70-80% con uency, and xed for 10 minutes by 4% paraformaldehyde to make the DNA and protein xed and cross-linked. After cross-linking, they were randomly fragmented into pieces of appropriate sizes using a sonicator, and centrifuged at 13000 rpm at 4°C for 5 minutes to collect supernatant. The supernatant was divided into two parts and incubated overnight at 4°C with rabbit anti-IgG (ab109489, 1:100, Abcam, Cambridge, UK) and target protein speci c antibodies Ap1 (1:1000, ab21981, Abcam), TET1 (1:2000, ab220867, Abcam). The endogenous DNA protein complex was precipitated by protein agarose/Sepharose, and the nonspeci c complex was washed. The DNA was decrosslinked overnight at 65°C and puri ed by phenol/chloroform. The enrichment of Ap1 in TET1 promoter and the enrichment of TET1 in Nrf2 promoter were examined.
Statistical analysis SPSS 22.0 statistical software (IBM Corp. Armonk, NY, USA) was used to process the data. All experiments were repeated 3 times independently. All the data were presented as mean ± standard deviation. The comparisons between two groups were done by the unpaired t test and the comparisons among multiple groups were conducted by the one-way or two-way analysis of variance (ANOVA), followed by Tukey's multiple comparison test. A probability value of P < 0.05 indicated the difference was statistically signi cant.
Tan Y., Cao H., Li Q, & Sun J. (2021). The role of transcription factor Ap1 in the activation of the Nrf2/ARE pathway through TET1 in diabetic nephropathy. Cell biology international, 45(8).
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7

Chromatin Immunoprecipitation and Quantification

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HMCs were cultured to 70-80% con uency, and xed for 10 minutes by 4% paraformaldehyde to make the DNA and protein xed and cross-linked. After cross-linking, they were randomly fragmented into pieces of appropriate sizes using a sonicator, and centrifuged at 13000 rpm at 4°C for 5 minutes to collect supernatant. The supernatant was divided into two parts and incubated overnight at 4°C with rabbit anti-IgG (ab109489, 1:100, Abcam, Cambridge, UK) and target protein speci c antibodies Ap1 (1:1000, ab21981, Abcam), TET1 (1:2000, ab220867, Abcam). The endogenous DNA protein complex was precipitated by protein agarose/Sepharose, and the nonspeci c complex was washed. The DNA was decrosslinked overnight at 65°C and puri ed by phenol/chloroform. The enrichment of Ap1 in TET1 promoter and the enrichment of TET1 in Nrf2 promoter were examined.
Statistical analysis SPSS 22.0 statistical software (IBM Corp. Armonk, NY, USA) was used to process the data. All experiments were repeated 3 times independently. All the data were presented as mean ± standard deviation. The comparisons between two groups were done by the unpaired t test and the comparisons among multiple groups were conducted by the one-way or two-way analysis of variance (ANOVA), followed by Tukey's multiple comparison test. A probability value of P < 0.05 indicated the difference was statistically signi cant.
Tan Y., Cao H., Li Q., & Jianjun S. (2020). The Role of Transcription Factor Ap1 in The Activation of Nrf2/ARE Pathway Through TET1 in Diabetic Nephropathy.
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