Epitope mapped
Disease related
Highly published

anti-Keratin Type II mouse monoclonal, AE3, purified

Cat. No. 61806

Available, delivery time 1-7 days

€235.00*

Key Features
  • Mouse monoclonal
  • Suitable for ICC/IF, IHC and WB
  • Reacts with bovine, chicken, human, monkey, mouse, rabbit and rat
  • Isotype: IgG1
Product description
Quantity 1 ml (200 µg/ml)
Antibody Type Monoclonal
Host Mouse
Isotype IgG1
Conjugate Unconjugated
Application ICC/IF, IHC, WB
Purification Affinity chromatography
Reactivity Bovine, Chicken, Human, Monkey, Mouse, 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 AE3
Immunogen Human epidermal keratin
Concentration 200 µg/ml
Formulation PBS pH 7.4 with 0.09% sodium azide
Note Centrifuge prior to opening
Applications
Tested applications Tested dilutions
Immunocytochemistry (ICC)/ Immunofluorescence (IF) Assay dependent
Immunohistochemistry (IHC) - frozen 1:200
Immunohistochemistry (IHC) - paraffin 1:200 (protease treatment and/or microwave treatment recommended)
Western Blot (WB) Assay dependent
Background

AE 3 represents an excellent marker for distinguishing carcinoma from non-epithelial tumors. Tumors specifically detected: all epithelia and their neoplasms.
Polypeptide reacting: Mr 53,000 to Mr 67,000 basic type II keratin polypeptides of human epithelial proteins (keratins - formerly also designated cytokeratins - nos. K1 - K8). Epitope has been mapped to aa 266-299 on alpha-helical rod domain (see Waseem et al., 2004).
Reactivity on cultured cell lines: MCF-7, A-431, RT 112.


Waseem A, Karsten U, Leigh IM, Purkis P, Waseem NH, Lane BE: Conformational changes in the rod domain of human keratin 8 following heterotypic association with keratin 18 and its implication for filament stability. Biochemistry 43, 1283-1295 (2004)

References/Publications (10)
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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.

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.
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.
  • 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.

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