Key Features
- Mouse monoclonal
- Suitable for ICC/IF, IHC on frozen and paraffin sections and WB
- Reacts with dog, hamster, human, mouse, pig, rabbit and rat
- Isotype: IgG1
Product description
Quantity | 1 ml |
---|---|
Antibody Type | Monoclonal |
Host | Mouse |
Isotype | IgG1 |
Conjugate | Unconjugated |
Application | ICC/IF, IHC, WB |
Purification | Hybridoma cell culture supernatant |
Reactivity | Dog, Hamster, Human, Mouse, Pig, Rabbit, Rat |
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 | RCK102 |
Immunogen | Lung cancer cell preparation |
Formulation | Contains 0.09% sodium azide |
UniprotID | Q7RTS7 (Human),Q922U2 (Mouse),A0A287BA49 (Pig),A0A286X8W5 (Guinea pig),A0A5F9C3E1 (Rabbit),Q6P6Q2 (Rat),Q6P6Q2 (Rat),P05787 (Human),P11679 (Mouse) |
Synonym | Keratin, type II cytoskeletal 74, Cytokeratin-74, CK-74, Keratin-5c, K5C, Keratin-74, K74, Type II inner root sheath-specific keratin-K6irs4, Type-II keratin Kb37, KRT74, K6IRS4, KB37, KRT5C, KRT6IRS4, Keratin, type II cytoskeletal 8, Cytokeratin-8, CK-8, Keratin-8, K8, Type-II keratin Kb8, KRT8, CYK8 |
Note | Centrifuge prior to opening |
Applications
Tested applications | Tested dilutions |
---|---|
Immunocytochemistry (ICC)/ Immunofluorescence (IF) | Assay dependent |
Immunohistochemistry (IHC) - frozen | 1:5-1:10 |
Immunohistochemistry (IHC) - paraffin | 1:5-1:10 (microwave treatment recommended) |
Western Blot (WB) | Assay dependent |
Background
Mab RCK 102, directed against the 58 kD and 52.2 kD keratin proteins (keratins K5 and K8; formerly also designated cytokeratins 5 and 8 in the Moll catalogue of keratins), reacts with all epithelial cell types. No reaction is observed in non-epithelial tissues. The antibody can be used for the discrimination of carcinomas from non-epithelial tumors such as lymphomas, sarcomas and neural tumors. RCK 102 is applicable on frozen and paraffin-embedded tissue. Formalin fixation may cause antigen denaturation or masking. Therefore, protease digestion is necessary prior to antibody application.
Positive control: Skin, cervix.
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.
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.
Isolation of positive area from tissue slices under the microscope, followed by extraction procedure to enrich for the cytoskeleton.
Positive tissue: outer root sheath of hair follicle, sweat gland epithelium on the foot pads, filiform papillae of tongue.
Positive tissue: outer root sheath of hair follicle, sweat gland epithelium on the foot pads, filiform papillae of tongue.
- Homogenization (e.g. with polytron) of tissue samples in buffer L (140 mM NaCl, 5 mM EDTA, 5 mM EGTA, 10 mM Tris-HCl pH7.6) supplemented with protease inhibitors; use 1 ml buffer for approx. 0.1 g tissue.
- To reduce viscosity by high DNA contents, benzonase treatment can be included (30 min at 37°C).
- Centrifugation for 10 min (max. speed, table top centrifuge).
- Resuspend pellet in "Triton extraction buffer" (buffer L plus 1% Triton X-100) using e.g. a dounce homogenizer.
- Centrifugation for 10 min (max. speed, table top centrifuge).
- Resuspend pellet in "high salt extraction buffer" (buffer L plus 1.5 M KCl) using e.g. a dounce homogenizer.
- Centrifugation for 10 min (max. speed, table top centrifuge).
- Resuspend pellet in "Triton extraction buffer" (buffer L plus 1% Triton X-100) using e.g. a dounce homogenizer.
- Centrifugation for 10 min (max. speed, table top centrifuge).
- Resuspend pellet in Laemmli buffer and boil (5 min) for SDS-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.