IHC analysis of human squamous cell carcinoma using anti-p16 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-p16 (Cat. No. 690074) was diluted in PBS (antibody concentration 2 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. The 20x picture was acquired using microscopy (courtesy of J.Hess, University Hospital Heidelberg).
Western blot analysis of HeLa lysate with anti-p16 antibody. Western blot analysis was performed on 20 µg HeLa lysate. Cells were lysed with RIPA buffer. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-p16 mouse monoclonal, DCS-50 (Cat. No. 690074) was diluted in blocking buffer (antibody concentration 1 µg/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG goat polyclonal, 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.
IHC analysis of human squamous cell carcinoma using anti-p16 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-p16 (Cat. No. 690074) was diluted in PBS (antibody concentration 2 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. The 20x picture was acquired using microscopy (courtesy of J.Hess, University Hospital Heidelberg).
Western blot analysis of HeLa lysate with anti-p16 antibody. Western blot analysis was performed on 20 µg HeLa lysate. Cells were lysed with RIPA buffer. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-p16 mouse monoclonal, DCS-50 (Cat. No. 690074) was diluted in blocking buffer (antibody concentration 1 µg/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG goat polyclonal, 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.
Key Features
- Purified, lyophilized
- Mouse monoclonal
- Suitable for ICC/IF, IHC and WB
- Reacts with human
- Isotype: IgG1
Product description
| Quantity | 50 µg |
|---|---|
| Antibody Type | Monoclonal |
| Host | Mouse |
| Isotype | IgG1 |
| Conjugate | Unconjugated |
| Application | ICC/IF, IHC, WB |
| Purification | Affinity chromatography |
| Reactivity | Human |
| 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 |
| Clone | DCS-50 |
| Immunogen | Recombinant human p16 protein |
| 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 | Q9UH64 (Human) |
| Synonym | Putative protein CDKN2A-DT, CDKN2A antisense RNA 1, CDKN2A antisense gene protein 1, Protein CDKN2A-AS1, Susceptibility protein NSG-x, CDKN2A-DT, C9orf53, CDKN2A-AS1 |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunocytochemistry (ICC)/ Immunofluorescence (IF) | Assay dependent |
| Immunohistochemistry (IHC) - paraffin | 1:10-1:50 (1-5 µg/ml; microwave treatment recommended) |
| Western Blot (WB) | 1:50-1:200 (0.25 µg/ml-1 µg/ml) |
Background
p16 (CDKN2A, p16Ink4A), is key regulator of the cell cycle and involved in cell cycle control and cellular senescence. It is a specific inhibitor for Cdk4 and Cdk6 and binds to the phosphorylated Cdk-cyclin complex. A disruption of this pathway is commonly observed in cancer.
p16 is lost in the majority of tumor cell lines and in most primary tumors. It is not expressed in melanoma. In carcinoma driven by an HPV (human papilloma virus) infection, p16 is often overexpressed.
The antibody is especially useful for immunoprecipitation.
The epitope was localized within the 15 aa residues of the C-terminus of p16 protein.
References/Publications (3)
Publication
Species
Application
Species
human
Application
ICC-IF
Species
human
Application
IHC (paraffin)
Species
human
Application
WB,ICC-IF
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FAQs
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.
PROGEN offers different formats for mouse monoclonal antibodies. Different antibodies may not be offered in each format, the following are all possible formats:
- Supernatant and supernatant concentrate: This format contains hybridoma cell culture supernatant. The antibody is not purified and the antibody concentration is not determined. The antibody concentration may vary from lot to lot. Therefore we recommend to titrate the optimal concentration for the application used for each new lot.
- Lyophilized, purified: This format contains purified antibody in lyophilized form. The reconstitution of this antibody is described in the datasheet. The buffer composition after reconstitution is also mentioned on the datasheet.
- Liquid, purified: This format contains purified antibody in liquid format. The concentration is mentioned on the datasheet.
- Prediluted, purified: This format contains purified antibody in liquid format. Most antibodies in this format are diluted to be ready-to-use for IHC with standard tissue. But some antibodies of this format need further dilution for IHC. This is mentioned on 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.
We offer bulk amounts starting from 1 mg for many of our mouse monoclonal or recombinant antibodies. Please contact us for further information or use the “bulk button” on the product page.
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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