anti-Vimentin guinea pig polyclonal, lyophilized, purified
Western blot analysis of HeLa lysate with anti-Vimentin antibody. Western blot analysis was performed on 10 µg wild type (WT) and 10 µg Vimentin knockout (KO) HeLa lysate. The PVDF membrane was blocked with 5% milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in blocking buffer (antibody concentration 0.02 µg/ml) and incubated for 1 h at RT. The secondary antibody anti-guninea pig, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
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IHC analysis of human head and neck squamous cell carcinoma using anti-Vimentin antibody. IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in PBS (antibody concentration 0.5 µg/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visable and with Haemalaun for a few minutes (courtesy of J. Hess, University Hospital Heidelberg).
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IHC analysis of murine kidney using anti-Vimentin antibody. IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in PBS (antibody concentration 1 µg/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visable and with Haemalaun for a few minutes (courtesy of J. Hess, University Hospital Heidelberg).
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Western blot analysis of human HeLa and mouse C2C12 lysate with anti-Vimentin antibody. Western blot analysis was performed on 20 µg HeLa and 18 µg C2C12 lysate. The PVDF membrane was blocked with 5% milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in blocking buffer (antibody concentration 0.02 µg/ml) and incubated for 1 h at RT. The secondary antibody anti-guinea pig, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
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IHC analysis of murine tongue using anti-Vimentin antibody. IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in PBS (antibody concentration 5 µg/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visable and with Haemalaun for a few minutes (courtesy of J. Hess, University Hospital Heidelberg).
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Western blot analysis of HeLa lysate with anti-Vimentin antibody. Western blot analysis was performed on 10 µg wild type (WT) and 10 µg Vimentin knockout (KO) HeLa lysate. The PVDF membrane was blocked with 5% milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in blocking buffer (antibody concentration 0.02 µg/ml) and incubated for 1 h at RT. The secondary antibody anti-guninea pig, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
View more
IHC analysis of human head and neck squamous cell carcinoma using anti-Vimentin antibody. IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in PBS (antibody concentration 0.5 µg/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visable and with Haemalaun for a few minutes (courtesy of J. Hess, University Hospital Heidelberg).
View more
IHC analysis of murine kidney using anti-Vimentin antibody. IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in PBS (antibody concentration 1 µg/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visable and with Haemalaun for a few minutes (courtesy of J. Hess, University Hospital Heidelberg).
View more
Western blot analysis of human HeLa and mouse C2C12 lysate with anti-Vimentin antibody. Western blot analysis was performed on 20 µg HeLa and 18 µg C2C12 lysate. The PVDF membrane was blocked with 5% milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in blocking buffer (antibody concentration 0.02 µg/ml) and incubated for 1 h at RT. The secondary antibody anti-guinea pig, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
View more
IHC analysis of murine tongue using anti-Vimentin antibody. IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-Vimentin guinea pig polyclonal (Cat. No. 610130) was diluted in PBS (antibody concentration 5 µg/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visable and with Haemalaun for a few minutes (courtesy of J. Hess, University Hospital Heidelberg).
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Key Features
- Guinea pig polyclonal
- Suitable for ELISA, IHC and WB
- Reacts with human and mouse
Product description
| Quantity | 50 µg |
|---|---|
| Antibody Type | Polyclonal |
| Host | Guinea pig |
| Conjugate | Unconjugated |
| Application | ELISA, IHC, WB |
| Purification | Affinity chromatography |
| Reactivity | Human, Mouse |
| Storage before reconstitution | 2-8°C until indicated expiry date |
| Storage after reconstitution | Up to 3 months at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles |
| Intended use | Research use only |
| Immunogen | Human Vimentin |
| Concentration | 50 µg/ml after reconstitution with 1 ml dist. water |
| Formulation | Lyophilized; reconstitute in 1 ml dist. water (final solution contains 0.09% sodium azide, 0.5% BSA in PBS buffer, pH 7.4) |
| UniprotID | P08670 (Human), P20152 (Mouse) |
| Synonym | Vimentin, VIM |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunohistochemistry (IHC) - paraffin | 1:10-1:100 (0.5-5 µg/ml, microwave treatment recommended) |
| ELISA | Assay dependent |
| Western Blot (WB) | 1:3,000 (0.02 µg/ml) |
Background
Specific detection of vimentin (57 kDa polypeptide). Tumors specifically detected: Sarcoma (including myosarcoma), lymphoma, melanoma.
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FAQs
- PVDF membranes show better results than nitrocellulose (higher capacity, allows for more stringent washing conditions in case of background problems).
- Use freshly prepared blocking solution (e.g. 5% nonfat dry milk, 0.05% Tween 20), block for at least 1 h at room temperature.
- Use the antibody in a higher dilution, but prolong incubation time and exposure time.
- Always use a fresh aliquot of the antibody.
- Do not repeatedly freeze the antibody (eventually centrifuge shortly after thawing to remove cryo-precipitates).
- Include an additional washing step.
You might also try more stringent wash conditions, e.g. add 0.5 M NaCl to the wash buffer. - Always use a fresh aliquot of secondary antibody.
- In case you use ECL most the guinea pig antibody should be diluted further in order to get rid of the background.
The concentration of unpurified supernatant, ascites, unpurified guinea pig serum and unpurified rabbit serum is not determined.
The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at support@progen.com.
The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at support@progen.com.
Most of our purified mouse antibodies contain 0.5% BSA as stabilizer. If BSA was added to the antibody solution, it is stated in the datasheet.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
Please find the solvent for reconstitution and the amount of liquid for reconstitution on the antibody datasheet. The buffer formulation after reconstitution is also mentioned in the datasheet.
Lyophilized antibodies can be stored at 2-8°C until expiration.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
The expiration date of our antibodies is indicated on the product label.
Most of our antibodies contain 0.09% sodium azide as preservative. If a preservative is added, it is mentioned in the datasheet.
The optimal antibody dilution for your specific protocol and application needs to be titrated in your lab with your equipment and sample. The optimal dilution may vary between protocols and samples. A good dilution for starting the titration is the dilution mentioned in the datasheet. If the sample needs a specific treatment (eg. Antigen retrieval for IHC on FFPE sections) this should also be mentioned on the antibody datasheet.
PROGEN antibodies are shipped at ambient temperature. The antibodies are stable at ambient temperature for the shipment period. Please store the antibodies as indicated in the datasheet upon arrival.
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