Human colon carcinoma (courtesy of L. Langbein)
CaCo colon carcinoma cells (courtesy of L. Langbein)
Human mamma carcinoma cells (courtesy of L. Langbein)
Human colon carcinoma (courtesy of L. Langbein)
CaCo colon carcinoma cells (courtesy of L. Langbein)
Human mamma carcinoma cells (courtesy of L. Langbein)
Key Features
- Guinea pig polyclonal
- Suitable for ICC/IF, IHC and WB
- Reacts with bovine, human, mouse and rabbit
Product description
| Quantity | 100 µl |
|---|---|
| Antibody Type | Polyclonal |
| Host | Guinea pig |
| Conjugate | Unconjugated |
| Application | ICC/IF, IHC, WB |
| Purification | Stabilized antiserum |
| Reactivity | Bovine, Human, Mouse, Rabbit |
| Storage | Short term at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles |
| Intended use | Research use only |
| Immunogen | Full length GST-fusion protein of human cingulin |
| Formulation | Contains 0.09% sodium azide and 0.5% BSA |
| UniprotID | P23805 (Bovine),Q9P2M7 (Human),P59242 (Mouse),B7NZD9 (Rabbit) |
| Synonym | Cingulin, CGN, KIAA1319 |
| Note | Centrifuge prior to opening |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunocytochemistry (ICC)/ Immunofluorescence (IF) | 1:100 (initial fixation with formaldehyde recommended for optimal staining) |
| Immunohistochemistry (IHC) - frozen | assay dependent |
| Immunohistochemistry (IHC) - paraffin | Assay dependent (microwave treatment recommended) |
| Western Blot (WB) | Assay dependent |
Background
The antibody reacts specifically with cingulin (a ca.150 kD polypeptide) present in tight junctions (zonula occludens) of polar epithelia (e.g. colon, duodenum, kidney, liver, pancreas) of transitional epithelia (e.g. bladder epithelium) and of stratified epithelia (e.g. skin, foot sole, tongue, oesophagus, vagina).
It is negative with endothelia of vessels and capillaries (e.g. in heart and brain).
Polypeptide reacting: Cingulin, a polypeptide of 126.5 kD (calculated from aa sequence data); apparent Mr 150 kD (after SDS-PAGE); pI 6.5 (very similar to that of symplekin, another junctional protein).
Reactivity on cultured cell lines: Carcinoma cell lines of human(e.g. MCF-7, Caco-2, PLC, A-431), bovine (MDBK, BMGE) and canine (MDCK) origin.
Positive control: total cell lysate of e.g. CaCo-2 cells.
References/Publications (2)
Publication
Species
Application
Species
human
Application
IHC (frozen)
Species
mouse
Application
IHC (paraffin)
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.
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.
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.
- Methanol/ acetone fixation: Immerse slide in precooled (-20°C) methanol for 5 min, immerse in precooled (-20°C) acetone for 30-60 sec, let specimen air dry before antibody incubation.
- Methanol/ acetone fixation plus detergent permeabilization: After methanol/ acetone fixation and air-drying dip slide either in a solution containing 0.1-0.2% Triton X-100 in PBS or in 0.1% saponin in PBS for 1-5 min at room temperature (enhances accessibility of many cytoskeletal antigens).
- Air-drying of the section.
- Block with the serum of the species in which the secondary antibody was raised for 30 min.
- Incubation with 1st antibody 1 h at RT in moist chamber.
- Wash 3x with PBS.
- Incubation with appropriate fluorescent secondary antibody, 30-60 min at RT.
- Wash 3x with PBS.
- Immerse shortly into ethanol.
- Let air dry.
- Cover with mounting medium.
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.
The concentration of specific antibody in our guinea pig serum is not determined.
In guinea pigs the antibody concentration in serum varies from 10 to 20 mg/ml; specific antibodies represent normally about 0.1-1% of total IgG. Total protein concentration varies from 40 to 65 mg/ml, with the main constituent (about 60%) being albumin.
In guinea pigs the antibody concentration in serum varies from 10 to 20 mg/ml; specific antibodies represent normally about 0.1-1% of total IgG. Total protein concentration varies from 40 to 65 mg/ml, with the main constituent (about 60%) being albumin.
Related Products
anti-Cingulin mouse monoclonal, 139.3.4, supernatant
€202.00*
Cat. No. 651122
- Mouse monoclonal
- Suitable for ICC/IF, IHC and WB
- Reacts with bovine, human and mouse
- Isotype: IgM