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anti-Desmocollin 3 mouse monoclonal, Dsc3-U114, lyophilized, purified

Key Features
  • 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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Category Protocol
Size 167.19 KB
Filetype pdf
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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.

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.

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:
  1. 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.
  2. 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. 
  3. Liquid, purified: This format contains purified antibody in liquid format. The concentration is mentioned on the datasheet. 
  4. 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.

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. 
  • 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).

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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