Species/Reactant: Homo sapiens (Human)
Applications: Immunocytochemistry-immunofluorescence
Image collected and cropped by CiteAb from the following publication, provided under a CC-BY licence.
Species/Reactant: Homo sapiens (Human)
Applications: Immunocytochemistry-immunofluorescence
Image collected and cropped by CiteAb from the following publication, provided under a CC-BY licence.
Species/Reactant: Homo sapiens (Human)
Applications: Immunocytochemistry-immunofluorescence
Image collected and cropped by CiteAb from the following publication, provided under a CC-BY licence.
Species/Reactant: Homo sapiens (Human)
Applications: Western Blotting
Image collected and cropped by CiteAb from the following publication, provided under a CC-BY licence.
Species/Reactant: Homo sapiens (Human)
Applications: Immunocytochemistry-immunofluorescence
Image collected and cropped by CiteAb from the following publication, provided under a CC-BY licence.
Species/Reactant: Homo sapiens (Human)
Applications: Immunocytochemistry-immunofluorescence
Image collected and cropped by CiteAb from the following publication, provided under a CC-BY licence.
Species/Reactant: Homo sapiens (Human)
Applications: Immunocytochemistry-immunofluorescence
Image collected and cropped by CiteAb from the following publication, provided under a CC-BY licence.
Species/Reactant: Homo sapiens (Human)
Applications: Western Blotting
Image collected and cropped by CiteAb from the following publication, provided under a CC-BY licence.
- Purified, lyophilized
- Mouse monoclonal
- Suitable for IHC and WB
- Reacts with human, mouse, rat
- Isotype: IgG1
Product description
| Quantity | 50 µg |
|---|---|
| Antibody Type | Monoclonal |
| Host | Mouse |
| Isotype | IgG1 |
| Conjugate | Unconjugated |
| Application | IHC, WB |
| Purification | Affinity chromatography |
| Reactivity | Human, Mouse, Rat |
| No reactivity | Bovine |
| 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 | Dsc3-U114 |
| Immunogen | Synthetic peptide corresponding to a sequence present on the extracellular anchor domain of human desmocollin 3 (VHGAPFYFSLPNTSPEISRLWSLTKVC) |
| 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 | Q14574 (Human), P55292 (Mouse), D3ZZI6 (Rat) |
| Synonym | Desmocollin-3, Cadherin family member 3, Desmocollin-4, HT-CP, DSC3, CDHF3, DSC4 |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunohistochemistry (IHC) - frozen | 1:10-1:100 (for better resolution preincubation (directly after fixation) with 0.05-0.2% Triton X-100, for 5-10 min, depending on tissue type, is recommended) |
| Immunohistochemistry (IHC) - paraffin | 1:10-1:100 (microwave treatment recommended) |
| Western Blot (WB) | Assay dependent |
Background
The mouse monoclonal antibody localizes Dsc3 present in
- All living epidermal layers
- In glandular duct cells
- In basal matrix cells and outer root sheath of hair follicles
- Basal and suprabasal cell layers of stratified epithelia (e.g. vagina, tongue, esophagus), basal layer of bladder urothelium and complex epithelium of trachea
- Thymic reticulum cells
All simple epithelia are negative. Intercalated disks of myocardium are
negative. Polypeptide reacting: 109 kDa and 100 kDa polypeptides (desmocollin 3 splice isoforms at pI 5.2) from human epidermal desmosomes.
Reactivity on cultured cell lines: most cultured cell lines derived from stratified squamous epithelia or squamous cell carcinomas are positive (e.g. A-431, HaCaT).
References/Publications (9)
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FAQs
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 supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
- 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.
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.
- As a suitable control tissue we suggest stratified epithelia, e.g. from vagina, tongue, esophagus (all basal and suprabasal cell layers react positively).
- For better resolution on frozen sections preincubation with 0.05-0.2% Triton X-100, for 5-10 min is recommended (directly after fixation with e.g. formaldehyde).
- 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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