Epitope mapped
Highly published
anti-Plakoglobin mouse monoclonal, PG 5.1, liquid, purified, sample
IHC of mouse tongue (courtesy of J.Heß, University Hospital Heidelberg)
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Western blot analysis of HeLa lysate and with anti-Plakoglobin antibody. Western blot analysis was performed on 20 µg 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-Plakoglobin mouse monoclonal, PG 5.1 (Cat. No. 690005) was diluted in blocking buffer (antibody concentration 25 ng/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG, HRP conjugate was also diluted in blocking buffer (antibody concentration 200 ng/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 of human oral mucosa (courtesy of J.Heß, University Hospital Heidelberg)
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IHC of rat tongue (courtesy of J.Heß, University Hospital Heidelberg)
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Western blot analysis of Ins-1 lysate and with anti-Plakoglobin antibody. Western blot analysis was performed on 20 µg rat Ins-1 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-Plakoglobin mouse monoclonal, PG 5.1 (Cat. No. 690005) was diluted in blocking buffer (antibody concentration 25 ng/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG, HRP conjugate was also diluted in blocking buffer (antibody concentration 200 ng/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 of mouse tongue (courtesy of J.Heß, University Hospital Heidelberg)
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Western blot analysis of HeLa lysate and with anti-Plakoglobin antibody. Western blot analysis was performed on 20 µg 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-Plakoglobin mouse monoclonal, PG 5.1 (Cat. No. 690005) was diluted in blocking buffer (antibody concentration 25 ng/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG, HRP conjugate was also diluted in blocking buffer (antibody concentration 200 ng/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 of human oral mucosa (courtesy of J.Heß, University Hospital Heidelberg)
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IHC of rat tongue (courtesy of J.Heß, University Hospital Heidelberg)
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Western blot analysis of Ins-1 lysate and with anti-Plakoglobin antibody. Western blot analysis was performed on 20 µg rat Ins-1 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-Plakoglobin mouse monoclonal, PG 5.1 (Cat. No. 690005) was diluted in blocking buffer (antibody concentration 25 ng/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG, HRP conjugate was also diluted in blocking buffer (antibody concentration 200 ng/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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Key Features
- Mouse monoclonal
- Suitable for ICC/IF, IHC and WB
- Reacts with bovine, dog, human, mouse, rat and zebrafish
- Isotype: IgG2b
Product description
| Quantity | 200 µl |
|---|---|
| Antibody Type | Monoclonal |
| Host | Mouse |
| Isotype | IgG2b |
| Conjugate | Unconjugated |
| Application | ICC/IF, IHC, WB |
| Purification | Affinity chromatography |
| Reactivity | Bovine, Dog, Human, Mouse, Rat, Zebrafish |
| Storage | Short term at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles |
| Intended use | Research use only |
| Clone | PG 5.1 |
| Immunogen | Plakoglobin, "band 5" protein from bovine snout epidermis |
| Concentration | 50 µg/ml (10 µg) |
| Formulation | PBS pH 7.4 with 0.09% sodium azide and 0.5% BSA |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunocytochemistry (ICC)/ Immunofluorescence (IF) | 1:5 (10 µg/ml) |
| Immunohistochemistry (IHC) - frozen | 1:25-1:100 (0.5-2 µg/ml) |
| Immunohistochemistry (IHC) - paraffin | 1:25-1:100 (0.5-2 µg/ml, microwave treatment recommended) |
| Western Blot (WB) | 1:500-1:2,000 (0.025-0.1 µg/ml) |
Background
PG 5.1 represents an excellent marker for all forms of intercellular adhering junctions, such as: desmosomes of epithelial and myocardial cells (incl. cultured cells); zonulae and fasciae adherentes of epithelia, endothelia of blood vessels and myocardial cells; adherens-type junctions (e.g. lens tissue, pigmented retinal cells, Sertoli cells of testis). The PG 5.1 epitope maps within the C-terminus at the extreme end of repeat 13 (aa 632-687) of plakoglobin. Polypeptide reacting: Mr 83,000 Plakoglobin, 'band 5' polypeptide of intercellular adhering junctions (identical to gamma-catenin).
Reactivity on cultured cell lines: cell cultures forming monolayers.
References/Publications (20)
Publication
Species
Application
Species
human
Application
WB
Species
mouse
Application
IHC (frozen), WB
Species
mouse
Application
WB,IHC (frozen)
Species
human
Application
WB,IHC
Species
human
Application
ICC-IF
Species
mouse
Application
WB
Species
mouse
Application
WB,IHC
Species
human
Application
IHC-IF (frozen)
Species
human,mouse,rat,bovine,eel,carp
Application
WB
Species
human
Application
WB,IP
Species
human
Application
ICC-IF
Species
human
Application
WB
Species
mouse
Application
ICC-IF
Species
Human, Zebrafish
Application
WB, ICC-IF, IP
Species
human
Application
IHC (frozen)
Species
human
Application
WB
Species
human
Application
WB
Species
human
Application
IHC (frozen),ICC-IF
Species
human
Application
IHC (frozen)
Species
human
Application
WB,ICC-IF,IEM
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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.
- Homogenization of tissue in PBS (supplemented with protease inhibitors).
- Centrifugation at 10E4 x g.
- Optional: treat pellet with DNase.
- Resuspend pellet in detergent buffer (e.g. 1% Triton X-100), stir for 1 h at 2-8°C.
- Centrifugation at 10E4 x g.
- Resuspend pellet in high salt buffer (with 1.5 M KCl), stir for 1 h at 2-8°C.
- Centrifugation at 10E4 x g.
- Resuspend pellet in low salt buffer (e.g. 50 mM NaCl), stir for 30 min at 2-8°C.
- Centrifugation at 10E4 x g: pellet contains raw cytoskeletal fraction.
- Transfer from a low concentration acrylamide gel (e.g. 8%).
- Reduce the methanol concentration of the transfer buffer (limit is 50% of the routine concentration 20% methanol).
- Use the wet transfer method and transfer at least 4 h (or better overnight) at 100-200 mA.
- In order to prevent the accumulation of gas bubbles, stir the transfer buffer continuously during the transfer (e.g. with a magnetic stirrer).
- If possible use a cytoskeletal preparation of your material to be analyzed (in the case of cultured cells additionally DNase treatment) rather than whole cell lysate.
- As a general rule we would like to recommend loading that much protein into one gel lane that the intermediate filament polypeptides (in the 40-60 kD range) can be readily detected in a control gel stained with Coomassie blue.
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