Anti-Vimentin Antibody Picoband®

Western blot analysis of Vimentin using anti-Vimentin antibody (PB9359).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: human U-87MG whole cell lysates,
Lane 3: human U20S whole cell lysates,
Lane 4: human PC-3 whole cell lysates,
Lane 5: human Hela whole cell lysates,
Lane 6: human THP-1 whole cell lysates,
Lane 7: human A549 whole cell lysates,
Lane 8: human MCF-7 whole cell lysates,
Lane 9: rat ovary tissue lysates,
Lane 10: rat lung tissue lysates,
Lane 11: mouse lung tissue lysates,
Lane 12: mouse NIH/3T3 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Vimentin antigen affinity purified polyclonal antibody (Catalog # PB9359) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Vimentin at approximately 54 kDa. The expected band size for Vimentin is at 54 kDa.

Western blot analysis of Vimentin using anti-Vimentin antibody (PB9359).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1-4: control group-human HTR8 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Vimentin antigen affinity purified polyclonal antibody (PB9359) at 1:2500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1 hour at RT. The signal is developed using an ECL Plus Western Blotting Substratewith ChemiDoc MP system. A specific band was detected for Vimentin at approximately 54 kDa. The expected band size for Vimentin is at 54 kDa.

IHC analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of mouse cardiac muscle tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of rat cardiac muscle tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of human lung adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of human invasive urothelial carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of Vimentin using anti-Vimentin antibody (PB9359).
Vimentin was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-Vimentin Antibody (PB9359) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

MiR-3619 induces p21, downregulates β-catenin and CDK2 expression and inhibits the EMT process. a p21, E-cadherin, and Cyclin D1 mRNA expression were detected by qRT-PCR. b , c The expression of proteins from potential miR-3619 targets is shown in a western blot of BCa cells from 3 days after transfection of miR-3619 or controls. d , e Immunofluorescence staining of β-catenin in 5637 and T24 cells. f N-cadherin, Vimentin, and Snail mRNA expression in 5637 and T24 cells were measured using qRT-PCR. g The N-cadherin, Vimentin, and Snail protein expression in 5637 and T24 cells were measured using western blot. * P < 0.05, ** P < 0.01 compared with the dsControl group
Index in PubMed under a CC BY license. PMID: 30237499

Generation of 3D-spheroid GMSCs. ( A ) 4 × 10 4 of GMSCs/well in 200 µl of complete stemgro culture medium was seeded into each well of ultra-low attachment round-shaped 96-U well plates and cultured for 48 h. H & E staining of cryosections of GMSC spheroids. Scale bar: 50 µm. ( B ) Immunocytochemistry showed the expression of a panel of MSC-associated markers CD29, CD73 and CD90 and extracellular components such as type I collagen (col-I), vimentin, fibronectin, and laminin in GMSC-derived spheroids. ( C ) GMSC spheroids were dissociated into single cells and stained with Annexin V-FITC and 7-AAD to detect early apoptosis and late apoptotic/necrotic cells by flow cytometric analysis. ( D ) Immunocytochemistry showed that less than 10% of cells inside GMSC spheroids were positive for the cleaved caspase-3. Cell nuclei were counter-stained by DAPI (blue). Scale bar: 50 µm. Data are representative of 3 independent experiments.
Index in PubMed under a CC BY license. PMID: 29700345

MiR-139 suppresses HCC migration and invasion. a - e Hep3B and SMMC7721 cells were transfected with 100 nM miR-139 mimic oligos or control oligos. The migration capacity of ( a ) Hep3B and ( b ) SMMC7721 cells were analyzed with the transwell migration assay. The invasion ability of ( c ) Hep3B and ( d ) SMMC7721 cells were measured with the matrigel transwell invasion assay. e The protein expression levels of E-cadherin, Vimentin and SNAIL1 were assayed with western blot in both Hep3B and SMMC7721 cells. f Hep3B and SMMC7721 cells were transfected with 100 nM miR-139 inhibitory or control oligos. The protein levels of E-cadherin, Vimentin and SNAIL1 were measured by western blot in both cell lines. All experiments were repeated 3 times. ** P < 0.01, *** P < 0.001
Index in PubMed under a CC BY license. PMID: 31046781

Down-regulating KPNA2 leads to decreased growth of HCC cells. Hep3B cells were transfected with non-targeting control oligo or with siRNA specifically targeting KPNA2 using lipofectamine 2000. a KPNA2 mRNA level was measured with qRT-PCR. b Western blot was used to assay the protein level of KPNA2. c Cell viability was measured using the MTT assay. d Cell apoptosis was measured with the Annxin V-FITC/Hoechst 33258 double staining flow cytometry. The cells were incubated for 72 h after transfection of the oligos. e colonogenic, ( f ) migratory and ( g ) invading capacities of cells were assayed as described above. h The protein levels of E-cadherin and Vimentin were measured using western blot. i The OS of HCC patients with low or high KPNA2 level was analyzed with Kaplan-Meier survival analysis and the curves were compared with the Log-rank test. The Q1–4 represent the 25% quantile of KPNA2 expression. Q1 represents the lowest 25%, Q2 represents 25–50%, Q3 is the 50–75% and Q4 is the 75–100%. All experiments were repeated 3 times. * P < 0.05, ** P < 0.01
Index in PubMed under a CC BY license. PMID: 31046781

LRRC4 inhibits collective cells invasion by down-regulating E-cadherin without EMT induction. (A) Phalloidin staining was performed in SKOV3 cells stably expressing LRRC4 through lentivirus infection. F-actin aggregates along control cell peripherals, while LRRC4 inhibites the accumulation and aggregation of F-actin (indicated with arrowheads). (B) The mRNA levels of EMT associated-genes, including E-cadherin, N-cadherin, slug, twist, ZEB1 and ZEB2, as assessed with RT-qPCR. (C) Western blotting analysis of E-cadherin and Vimentin protein in SKOV3 cells transiently transfected with different doses of LRRC4 expression vector (0.5, 1, and 2 μg). (D,E) E-cadherin and LRRC4 protein expression and localization as detected with immunohistochemistry (IHC) in human normal ovarian epithelial cells ( D , n = 5), human HGSC ( E , n = 17) and ascites of ovarian cancer patients. Scale bars: the left is 50 μm while the right is 20 μm. The area in the red boxes to the right is magnified. (F) E-cadherin and LRRC4 in EOC ascitic tissue as stained with IHC. Scale bars: 20 μm. (G) Expression of E-cadherin, Vimentin and Pan-cadherin in intraperitoneal xenografted primary tumor tissues from the mouse model was examined by the use of IHC in. Scale bars: 20 μm and 50 μm. * p < 0.05, ** p < 0.01.
Index in PubMed under a CC BY license. PMID: 32117780

Key characteristics of the immortalized HP-1 human PSC cell line. Phase contrast microscopy in HP-1 cells after seeding for 12 h ( A ); Double-immunofluorescence labelling using antibodies for SV40 antigen and GFAP, green, localization of SV40 antigen, red, localization of GFAP (B); Immunofluorescence showing the presence of α-SMA ( C ); Double-immunofluorescence labelling for vimentin and desmin. Green, localization of vimentin (D), red, localization of desmin (E), yellow, colocalization of vimentin and desmin (F). (Original magnification × 200 in A ; × 400 in B-F ).
Index in PubMed under a CC BY license. PMID: 31929747

ZFPM2-AS1 promoted cell metastatic properties by affecting EMT in HCC cells. A , B The migration abilities were evaluated in HCC cells infected with sh-ZFPM2-AS1-1 or sh-ZFPM2-AS1-2 lentivirus, by wound-healing assays. Scale bar = 50 μm. C Transwell assays detected the invasion capacities of HCC cells after ZFPM2-AS1-2 depletion. Scale bar = 50 μm. D Western blot examined the protein levels of N-cadherin and vimentin in HCC cells. Data are presented as the mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01.
Index in PubMed under a CC BY license. PMID: 33414427

The characterization of PDLCs. Primary culture of PDLCs at the Day 7 (×100) ( a ); Morphology of PDLCs at Passage 1 (× 100) ( b ); Immunohistochemistry staining results of PDLCs, positive for vimentin (× 100) ( c ) and negative for cytokeratin (× 100) ( d )
Index in PubMed under a CC BY license. PMID: 33485352

LPC inhibits pulmonary metastasis of CT26 colon cancer. a Schematic view of the experimental procedures of CT26 pulmonary metastatic mouse model. b , c Image and corresponding H&E staining of lung tissue. d Percentage change of body weight. e – g Lung weight, the number of lung tumor nodule, and metastasis rate ( n = 6 mice). h Protein expression of PCNA, vimentin and E-cadherin in lungs ( n = 3 mice). Data were presented as mean ± SEM, * p < 0.05, ** p < 0.01.
Index in PubMed under a CC BY license. PMID: 36774343

Western blot results showing the expression levels of A549 cells treated with different concentrations of GLXB-D drug serum and TGF-β1 for 24 h . (A): The protein expression straps of E-cadherin and Vimentin; (B): The grey value ratio of expressions of E-cadherin and Vimentin. (1) control group; (2) 5ng/mL TGF-β1 induced group; (3) 5% GLXB-D drug serum group; (4) 10% GLXB-D drug serum group; (5) 15% GLXB-D drug serum group.
Index in De Gruyter Brill under a CC BY license. DOI: 10.1515/chem-2018-0042