Human/Mouse/Rat Vimentin Antibody

Detection of Human, Mouse, and Rat Vimentin by Western Blot. Western blot shows lysates of K562 human chronic myelogenous leukemia cell line, MEF mouse embryonic feeder cells, RAW 264.7 mouse monocyte/macrophage cell line, NR8383 rat alveolar macrophage cell line, and Rat‑2 rat embryonic fibroblast cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human/Mouse/Rat Vimentin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2105) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for Vimentin at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Vimentin in Human Skin. Vimentin was detected in immersion fixed paraffin-embedded sections of human skin using 10 µg/mL Goat Anti-Human/Mouse/Rat Vimentin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2105) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Vimentin in HeLa Human Cell Line. Vimentin was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human/Mouse/Rat Vimentin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2105) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to intermediate filaments in cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human Vimentin by Simple Western^TM. Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line and Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Vimentin at approximately 59 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse/Rat Vimentin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2105) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Vimentin Specificity is Shown by Immunocytochemistry in Knockout Cell Line. Vimentin was detected in immersion fixed K562 human chronic myelogenous leukemia cell line but is not detected in Vimentin knockout (KO) K562 Human Cell Line cell line using Goat Anti-Human/Mouse/Rat Vimentin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2105) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL003) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our Fluorescent ICC Staining of Non-adherent Cells protocol.

Western Blot Shows Human Vimentin Specificity by Using Knockout Cell Line. Western blot shows lysates of K562 human chronic myelogenous leukemia parental cell line and Vimentin knockout K562 cell line (KO). PVDF membrane was probed with 2 µg/mL of Goat Anti-Human/Mouse/Rat Vimentin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2105) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Vimentin at approximately 55 kDa (as indicated) in the parental K562 cell line, but is not detectable in knockout K562 cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of Human Vimentin by Western Blot TGF-beta 1, -beta 2 and -beta 3 reduce lymphatic marker expression in LECs.Primary human LECs were treated with 10, 20 or 30 ng/ml TGF-beta 1, -beta 2 and -beta 3 for 72 hours (a) or 100 hours (b). Untreated cells served as a control. Lysates were prepared and analysed by Western blot using antibodies specific for Lyve-1, Prox-1, VEGFR-3 or vimentin. Vinculin served as loading control. The experiment was performed twice with equivalent results. For densitometry evaluation, protein bands were analysed using the software ImageJ. Bands for the Prox-1, Lyve-1, vimentin and VEGFR-3 proteins were normalized to the corresponding loading control and are displayed as the expression level relative to the untreated control samples. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0162221), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Vimentin by Western Blot Extracellular vesicles impact multiple signaling elements of the glioma stem cell program.a Mass spectrometry quantification (n = 3) for the presence of MMPs in HUVEC-EVs (EEVs), proneural-GSC157 cells, GSC157 cells treated with CMO and GSC157 cells treated with HUVEC-CM (CME). Comparisons are made relative to untreated GSC157 cells (b, c). MMP activity of GSC157 EVs, HUVEC-EVs and HBEC5i EVs over 1 h (n = 3; b) and cumulatively (n = 3; c). d Cumulative MMP activity for GSC157 cells, GSC157 cells treated with their own EVs (OEVs), HBEC5i EVs and HUVEC-EVs over 1 h (n = 3). e Cumulative MMP activity of GSC157 cells treated with HBEC5i-EVs and either DMSO control or MMP inhibitor, BB94 (n = 3). f Expression of NICD following treatment of proneural-GSC157 cells with OEVs or EEVs in the absence or presence of MMP inhibitors, AG3340 and BB94. g DAVID analysis of the top pathways enriched in proneural-GSC157 cells treated with CME (HUVEC-conditioned media). h Expression of proneural-(NICD) and mesenchymal-(VIM) hallmarks after pharmacological inhibition of: NF kappa B pathway (Bay11-7082), Wnt pathway (LGK974) and TGF beta pathway (LY2157299). i Immunocytochemistry and quantification of phospho-P65 (p-P65) after treatment of GSC157 cells with EEVs (HUVEC-EVs). Densitometry quantifications of p-P65 (j) and total P65 (k) in GSC157 cells treated with HBEC5i-EVs (EEVs) relative to OEVs (n = 3). l Expression of activated (phospho) and total p65 in the presence of Bay11-7082 in GSC157s. m Expression of activated (phospho) and total p65 in the presence of AG3340 and BB94 in GSC157 cells exposed to EEVs. n Quantification of wound healing assay for OEV, EEV or EEVs+Bay 11-7082 treated proneural-GSC157 cells. o Schematic of the different donor EVs (OEVs and EEVs) competing for being taken up by proneural-GSC157. p Either 5,000 or 50,000 GSC157 cells exposed to a fixed amount of 30μg of HBEC5i-EVs (EEVs). q Expression of NICD in the presence of OEVs, HUVEC-EVs (EEVs), OEV:EEV (1:1) and OEV:EEV (2:1). NICD Notch intracellular domain, CMO own conditioned meida, CME endothelial conditioned media, EV extracellular vesicles, OEVs own EVs, EEVs endothelial cell derived EVs, MMP matrix metaloprotinase; HUVEC human umbilical vein endothelial cells, HBEC5i immortalized human brain brain endothelial cells. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/36123372), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Vimentin by Western Blot NRF1 and/or E2 re-programming contributed to the overexpression of pluripotency and epithelial-mesenchymal transition (EMT) markers and differentiation of tumor initiating breast cancer cells to other-types of cells. Experimental conditions were the same as Figure 1. (A) Immunofluorescent and flow cytometry detections of MCF10A (T) tumor initiating pluripotent stem cells markers (SOX2, Oct4, and Nanog) in transformed breast epithelial cells of MCF10A [MCF-10A (T)] overexpressing vector (control) or NRF1 treated with DMSO and E2. (B) Immunofluorescent and flow cytometry detections of pluripotent stem cells markers (SOX2 and Oct4) in transformed breast epithelial cells of MCF10A [MCF-10A (T)] overexpressing vector (control) or NRF1 treated with DMSO and E2. (C) Immunofluorescent and Western detections of the expression of EMT markers E-cadherin, N-cadherin, and vimentin in transformed breast epithelial cells of MCF10A [MCF-10A (T)] overexpressing vector (control) or NRF1 treated with DMSO and E2. (D) Differentiation of NRF1 tumor initiating stem cells to chondrocytes, neurons, and smooth muscle cells compared to the vector. The vector did not show any differentiation to other cells. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30486409), licensed under a CC-BY license. Not internally tested by R&D Systems.