rabbit anti human β actin polyclonal antibody  (Proteintech)

Journal: Kidney360

Article Title: Serum AKR1A1 Levels Predict eGFR Decline Rate in Black Americans with Type 2 Diabetes

doi: 10.34067/KID.0000000966

Figure Lengend Snippet: Human immortalized kidney PTC line AKR1A1 KO and its effect on cell viability in high glucose and high palmitate environments. (A) AKR1A1 protein levels were depleted in immortalized AKR1A1 KO PTCs, compared with WT cells. Relative AKR1A1 protein levels were assessed by Western blot. Eight micrograms of cell lysate protein (lanes 2–5) were fractionated by 4%–20% SDS-PAGE and transferred to nitrocellulose membrane. The membrane was probed with Santa Cruz mouse anti- AKR1A1 antibody and Bioss rabbit anti- β -actin antibody. Lanes 1 and 6: mol wt marker; lane 2: WT PTC line; lane 3: AKR1A1 KO cell line clone 1; lane 4: AKR1A1 KO cell line clone 2; lane 5: AKR1A1 KO cell line clone 3. AKR1A1 KO PT clone 1 cells were used in the subsequent cell viability assays to represent KO cells. (B) AKR1A1 KO PTC viability was lower than WT PTCs and higher glucose concentrations did not impact cell viability in WT or AKR1A1 KO PTCs. G100, G500, and G1000 represent cells treated with glucose at 100, 500, and 1000 mg/dl, respectively, for 48 hours. Pair-wise P values were adjusted for Bonferroni correction. (C) AKR1A1 KO PTC viability was lower than WT PTCs, and higher palmitate concentration (0.5 mM) worsened cell viability in KO PTCs compared to KO PTCs treated without additional palmitate or with low concentration (0.5 mM) of palmitate. P0.00, P0.25, and P0.50 represent cells treated with additional palmitate at final concentrations 0.00, 0.25, and 0.50 mM, respectively, in growth media for 48 hours. Cell relative viability of cells was estimated as a percentage of baseline WT controls, where the mean of WT control repeats was set as 1 (100%). Pair-wise P values were adjusted for Bonferroni correction. KO, knockout; PT, proximal tubule; PTC, proximal tubule cell; WT, wild type.

Article Snippet: The blot was incubated overnight at 4°C with primary mouse monoclonal anti-human AKR1A1 (1:500; Santa Cruz, sc-271723) and rabbit polyclonal anti-human β -actin (1:3000; Bioss Antibodies, bs-0061R).

Techniques: Western Blot, SDS Page, Membrane, Marker, Concentration Assay, Control, Knock-Out

Journal: bioRxiv

Article Title: Metabolomic Profiling and Characterization of a Novel 3D Culture System for Studying Chondrocyte Mechanotransduction

doi: 10.1101/2024.06.10.598340

Figure Lengend Snippet: Western blot analysis of primary human and bovine chondrocytes. Monolayer cell lysates from (A) primary human chondrocytes and (B) bovine chondrocytes were probed with antibody to Collagen VI. Lanes 1 and 4 represent cell lysates from day 0, lanes 2 and 5 represent cell lysates from monolayers cultured for 7 days without ascorbate (Asc). Cell lysates from monolayers cultured with ascorbate (Asc) for 7 days are shown in lanes 3 and 6. β-actin was used as loading control.

Article Snippet: Loading controls were detected by running equal amounts of each lysate on a separate gel and probed with Rabbit Polyclonal Anti-Human, Anti-Bovine β actin (Rockland Immunochemicals).

Techniques: Western Blot, Cell Culture, Control

Journal: Cell reports

Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

doi: 10.1016/j.celrep.2023.112646

Figure Lengend Snippet: (A) Representative immunoblots of 3 independent studies of ITG proteins in Nsi and DAP5-silenced MB-231 cells. Dox induction of shRNAs for 4 days followed by immunoblot analysis of equal protein amounts. (B) Representative immunoblots of 3 independent studies of EMT biomarker protein levels in Nsi and DAP5-silenced 4T1 and MB-231 cells carried out as in (A). (C) Representative immunoblots of 3 independent studies of eIF4E and eIF2α proteins in Nsi and DAP5-silenced MB-231 cells carried out as in (A). (D) Representative immunoblots of 3 independent studies to test requirement for eIF3d cap binding activity for DAP5-dependent mRNAs in MB-231 cells. Cells were small interfering RNA (siRNA) silenced for eIF3d for 1 day and transfected with vectors expressing WT, α5, or α11 cap binding mutants of eIF3d for 2 days, and equal protein amounts analyzed using immunoblot. (E) Matrigel Transwell invasion assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Mean number of invading cells/field with SEM from 3 replicates for each cell line. See also . (F) Cell migration wound healing assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Time 0 represents 100% wound separation of the cell layer. Mean of percentage of migrated cell surface areas (percentage closure) with SEM from 3 replicates for each cell line. See also . (G) Induction of apoptosis by cell detachment (anoikis) determined in 4T1 and MB-231 cells silenced for 24 and 48 h for Nsi, DAP5, or eIF4GI, comparing cells maintained on adherent or ultra- low-adherence plates. Percentage cell apoptosis determined by annexin V-fluorescein isothiocyanate (FITC) staining and flow cytometry, quantified using FlowJo software. Mean with SEM of 3 independent studies per cell line. (H) Representative immunoblot of 3 independent studies of DAP5, eIF4G1, eIF3d, and control β-actin protein levels in Nsi or DAP5-silenced 4T1 and MB-231 cells, silenced for 2 days with Dox-inducible shRNAs. Data are represented as mean ± SEM. n.s., not significant. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by unpaired parametric two-tailed t test.

Article Snippet: Rabbit polyclonal anti-human/mouse β-actin antibody , Cell Signaling Technology , Cat# 4967/RRID:AB_330288.

Techniques: Western Blot, Biomarker Discovery, Binding Assay, Activity Assay, Small Interfering RNA, Transfection, Expressing, Migration, Staining, Flow Cytometry, Software, Control, Two Tailed Test

Journal: Cell reports

Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

doi: 10.1016/j.celrep.2023.112646

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-human/mouse β-actin antibody , Cell Signaling Technology , Cat# 4967/RRID:AB_330288.

Techniques: Recombinant, Staining, Protease Inhibitor, Virus, Reverse Transcription, SYBR Green Assay, Bicinchoninic Acid Protein Assay, MTT Assay, Software, Imaging