Vimentin | HUABIO EM0401 product information

product summary

company name :

HUABIO

product type :

antibody

product name :

Vimentin

catalog :

EM0401

quantity :

100μl

price :

360.00 USD

clonality :

monoclonal

host :

mouse

conjugate :

nonconjugated

clone name :

D4-B11

reactivity :

human, mouse, rat

application :

western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section, immunohistochemistry - frozen section

more info or order :

HUABIO product webpage

image

image 1 :

Western blot analysis of Vimentin with anti-Vimentin antibody [D4-B11] (EM0401) at 1/5,000 dilution. Lane 1: Wild-type Hela whole cell lysate (20 µg). Lane 2: Vimentin knockout Hela whole cell lysate (20 µg). Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (EM0401, 1/5,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

image 2 :

Western blot analysis of Vimentin on different lysates with Mouse anti-Vimentin antibody (EM0401) at 1/5,000 dilution. Lane 1: 293T cell lysate Lane 2: HeLa cell lysate Lane 3: Jurkat cell lysate Lane 4: C2C12 cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: C6 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 54 kDa Observed band size: 54 kDa Exposure time:15 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM0401) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

image 3 :

Immunofluorescence analysis of paraffin-embedded human kidney tissue labeling Vimentin (EM0401) and NPHS2 (ET7107-34). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Vimentin (EM0401, green) at 1/400 dilution and NPHS2 (ET7107-34, red) at 1/100 dilution overnight at 4 ℃, washed with PBS. iFluor™ 488 conjugate-Goat anti-Mouse IgG (HA1125) and iFluor™ 594 conjugate-Goat anti-Rabbit IgG (HA1122) were used as the secondary antibodies at 1/1,000 dilution. DAPI was used as nuclear counterstain.

product information

SKU :

EM0401

Target name :

Vimentin

Species reactivity :

Human,Mouse,Rat

Applications :

WB,IF-Cell,IHC-P,FC,IF-Tissue,IHC-Fr,IP

Conjugate :

Non-conjugated

Immunogen :

Synthetic peptide within C-terminal human Vimentin.

Uniprot id :

P08670>SwissProt: P08670 Human;SwissProt: P20152 Mouse;SwissProt: P31000 Rat

Host :

Mouse

Clone number :

D4-B11

Isotype :

IgG1

Size :

100μl

List Price :

360.00 USD

Storage Buffer :

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Form :

Liquid

Storage Instruction :

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Purity :

Protein A affinity purified.

Product type :

Mouse monoclonal Antibody

Positive control :

HeLa cell lysate, C2C12 cell lysate, C6 cell lysate, NIH/3T3 cell lysate, Mouse embryonic stem cell lysate, Hela, human kidney tissue, human colon carcinoma tissue, human stomach carcinoma tissue, human tonsil tissue, human skin tissue, human liver tissue, human appendix tissue, human endometrium tissue.

Molecular wt :

Predicted band size: 54 kDa

Subcellular location :

Cytoplasm

Concentration :

2 mg/mL.

Recommended dilutions :

WB: 1:5,000-1:10,000 ;IF-Cell: 1:200 ;IHC-P: 1:500-1:1,000 ;FC: 1:500-1:1,000 ;IF-Tissue: 1:200-1:500 ;IHC-Fr: 1:500-1:1,000 ;IP: Use at an assay dependent concentration

Advanced Validation :

Knockout (KO)

Pic img4 :

https://storage.huabio.cn/huabio/productImg/EM0401_4.jpg

Pic legend4 :

Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling Vimentin (EM0401) at 1/200 dilution and Cytokeratin 5 (ET1610-43) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Vimentin (EM0401, green) at 1/200 dilution and Cytokeratin 5 (ET1610-43, red) at 1/200 dilution at +4℃ overnight, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Mouse IgG and Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG were used as the secondary antibodies at 1/1,000 dilution. DAPI was used as nuclear counterstain.

Pic img5 :

https://storage.huabio.cn/huabio/productImg/EM0401_5.jpg

Pic legend5 :

Immunofluorescence analysis of paraffin-embedded human skin tissue labeling Vimentin (EM0401) at 1/200 dilution and Cytokeratin 5 (ET1610-43) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Vimentin (EM0401, green) at 1/200 dilution and Cytokeratin 5 (ET1610-43, red) at 1/200 dilution at +4℃ overnight, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Mouse IgG and Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG were used as the secondary antibodies at 1/1,000 dilution. DAPI was used as nuclear counterstain.

Pic img6 :

https://storage.huabio.cn/huabio/productImg/EM0401_6.jpg

Pic legend6 :

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Vimentin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0401, 1/800) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img7 :

https://storage.huabio.cn/huabio/productImg/EM0401_7.jpg

Pic legend7 :

Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue with Mouse anti-Vimentin antibody (EM0401) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0401) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img8 :

https://storage.huabio.cn/huabio/productImg/EM0401_8.jpg

Pic legend8 :

Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-Vimentin antibody (EM0401) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0401) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img9 :

https://storage.huabio.cn/huabio/productImg/EM0401_9.jpg

Pic legend9 :

Immunohistochemical analysis of paraffin-embedded human appendix tissue with Mouse anti-Vimentin antibody (EM0401) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0401) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img10 :

https://storage.huabio.cn/huabio/productImg/EM0401_10.jpg

Pic legend10 :

Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-Vimentin antibody (EM0401) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0401) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img11 :

https://storage.huabio.cn/huabio/productImg/EM0401_11.jpg

Pic legend11 :

Immunohistochemical analysis of paraffin-embedded human endometrium tissue with Mouse anti-Vimentin antibody (EM0401) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0401) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img12 :

https://storage.huabio.cn/huabio/productImg/EM0401_12.jpg

Pic legend12 :

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-Vimentin antibody (EM0401) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0401) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img13 :

https://storage.huabio.cn/huabio/productImg/EM0401_13.jpg

Pic legend13 :

Immunohistochemical analysis of paraffin-embedded rat colon tissue with Mouse anti-Vimentin antibody (EM0401) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0401) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img14 :

https://storage.huabio.cn/huabio/productImg/EM0401_14.jpg

Pic legend14 :

Vimentin was immunoprecipitated from 0.2 mg HeLa cell lysate with EM0401 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using EM0401 at 1/1,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: EM0401 IP in HeLa cell lysate Lane 3: Mouse IgG instead of EM0401 in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes; ECL: K1801

Pic img15 :

https://storage.huabio.cn/huabio/productImg/EM0401_15.jpg

Pic legend15 :

Application: IHC-Fr Species: Mouse Site: Colon Sample: Frozen section Antibody concentration: 1/500 Antigen retrieval: Not required

Pic img16 :

https://storage.huabio.cn/huabio/productImg/EM0401_16.jpg

Pic legend16 :

Immunocytochemistry analysis of HeLa cells labeling Vimentin with Mouse anti-Vimentin antibody (EM0401) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Vimentin antibody (EM0401) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

Pic img17 :

https://storage.huabio.cn/huabio/productImg/EM0401_17.jpg

Pic legend17 :

Immunocytochemistry analysis of C2C12 cells labeling Vimentin with Mouse anti-Vimentin antibody (EM0401) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Vimentin antibody (EM0401) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

Pic img18 :

https://storage.huabio.cn/huabio/productImg/EM0401_18.jpg

Pic legend18 :

Immunocytochemistry analysis of C6 cells labeling Vimentin with Mouse anti-Vimentin antibody (EM0401) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Vimentin antibody (EM0401) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

Pic img19 :

https://storage.huabio.cn/huabio/productImg/EM0401_19.jpg

Pic legend19 :

Flow cytometric analysis of Hela cells labeling Vimentin. Cells were fixed and permeabilized.Then stained with the primary antibody (EM0401, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

more info or order :

HUABIO product webpage