Vimentin Recombinant Rabbit Monoclonal Antibody [PDH0-01]
cat.: HA721174
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC, IF-Tissue, mIHC
Clonality: Monoclonal
Clone number: PDH0-01
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1.025ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 54 kDa
Isotype: IgG
Immunogen: Synthetic peptide within C-terminal human Vimentin.
Positive control: L6 cell lysate, C2C12 cell lysate, Hela cell lysate, A549 cell lysate, human liver tissue, NIH/3T3 cell lysate, C6 cell lysate, human appendix tissue, human kidney tissue, human stomach carcinoma tissue, human endometrium tissue, HeLa, NIH/3T3, human skin tissue, mouse lung tissue.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
  IF-Tissue
  mIHC
1:5,000
1:500-1:2,000
1:200
1:500-1:1,000
1:200
1:10,000
Uniprot #: SwissProt: P08670 Human | P20152 Mouse | P31000 Rat
Alternative names: CTRCT30 Epididymis luminal protein 113 FLJ36605 HEL113 VIM VIME_HUMAN Vimentin
Images
HA721174_1.jpg Fig1: Western blot analysis of Vimentin on different lysates with Rabbit anti-Vimentin antibody (HA721174) at 1/5,000 dilution.

Lane 1: L6 cell lysate
Lane 2: C2C12 cell lysate
Lane 3: HeLa cell lysate
Lane 4: A549 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 57 kDa

Exposure time: 2 minutes;

10% 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 (HA721174) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
HA721174_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Vimentin antibody (HA721174) 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 (HA721174) 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.
HA721174_3.jpg Fig3: Western blot analysis of Vimentin on different lysates with Rabbit anti-Vimentin antibody (HA721174) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: C6 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 54 kDa

Exposure time: 7 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 (HA721174) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
HA721174_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human appendix tissue with Rabbit anti-Vimentin antibody (HA721174) 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 (HA721174) 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.
HA721174_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Vimentin antibody (HA721174) 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 (HA721174) 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.
HA721174_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue with Rabbit anti-Vimentin antibody (HA721174) 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 (HA721174) 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.
HA721174_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human endometrium tissue with Rabbit anti-Vimentin antibody (HA721174) 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 (HA721174) 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.
HA721174_8.jpg Fig8: Immunocytochemistry analysis of HeLa cells labeling Vimentin with Rabbit anti-Vimentin antibody (HA721174) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Vimentin antibody (HA721174) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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.
HA721174_9.jpg Fig9: Immunocytochemistry analysis of NIH/3T3 cells labeling Vimentin with Rabbit anti-Vimentin antibody (HA721174) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Vimentin antibody (HA721174) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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.
HA721174_10.jpg Fig10: Immunofluorescence analysis of paraffin-embedded human kidney tissue labeling Vimentin with Rabbit anti-Vimentin antibody (HA721174) 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 antibody (HA721174, green) at 1/200 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HA721174_11.jpg Fig11: Immunofluorescence analysis of paraffin-embedded human skin tissue labeling Vimentin with Rabbit anti-Vimentin antibody (HA721174) 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 antibody (HA721174, green) at 1/200 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HA721174_12.jpg Fig12: Immunofluorescence analysis of paraffin-embedded human appendix tissue labeling Vimentin with Rabbit anti-Vimentin antibody (HA721174) 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 antibody (HA721174, green) at 1/200 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HA721174_13.jpg Fig13: Flow cytometric analysis of Hela cells labeling Vimentin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721174, 1ug/ml) (red) compared with Rabbit IgG 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-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA721174_14.jpg Fig14: mIHC analysis of human skin tissue (Formalin/PFA-fixed paraffin-embedded sections) with Rabbit anti-Vimentin antibody (HA721174) at 1/10,000 dilution. The immunostaining was performed with the IRISKit® HyperView mTSA Kit (MH900206). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
HA721174_15.jpg Fig15: mIHC analysis of mouse lung tissue (Formalin/PFA-fixed paraffin-embedded sections) with Rabbit anti-Vimentin antibody (HA721174) at 1/10,000 dilution. The immunostaining was performed with the IRISKit® HyperView mTSA Kit (MH900206). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
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