Highly published

anti-Keratin K5 guinea pig polyclonal, serum

Cat. No: GP-CK5

Available, delivery time 1-7 days

$365.00*

Key Features
  • Guinea pig polyclonal
  • Suitable for IHC and WB
  • Reacts with bovine, human and mouse
Product description
Quantity 100 µl
Antibody Type Polyclonal
Host Guinea pig
Conjugate Unconjugated
Application IHC, WB
Purification Stabilized antiserum
Reactivity Bovine, Human, Mouse
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 Recombinant human keratin K5
Formulation Contains 0.09% sodium azide and 0.5% BSA
UniprotID Q5XQN5 (Bovine), Q7RTS7 (Human), Q922U2 (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
Note Centrifuge prior to opening
Applications
Tested applications Tested dilutions
Immunohistochemistry (IHC) - frozen 1:100
Immunohistochemistry (IHC) - paraffin 1:50 (microwave treatment recommended)
Western Blot (WB) 1:1,500-1:5,000
Background

Basic keratin K5 (Mr 58,000; formerly also designated cytokeratin 5), expressed in basal and first suprabasal layers of epidermis. Highly specific staining pattern in human and murine skin sections.
Tumors Specifically Detected: The antiserum reacts with keratin K5, expressed in the basal cells of the larynx, esophagus, trachea, bladder, cervix, vagina, breast acini, skin and sweat glands.
In several studies the correlation between the expression of keratin K5 in different types of carcinomas (putatively derived from basal epithelial cells) and prognosis has been discussed (see publications).


References/Publications (8)
Publication
Species
Application
Downloads
File
Category
Size
Filetype
Category Protocol
Size 917.17 KB
Filetype pdf
Category Protocol
Size 919.99 KB
Filetype pdf
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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.

Colon
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.

  • Esophagus
  • Lacatating mammary gland
  • Urinary bladder
  • Sweet glands

  1. 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.
  2. 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.
  • Mild fixation with PFA (e.g. 2% PFA) is recommended.
  • Preincubation with 0.1-0.2% Triton-X-100 in PBS prior to fixation is optional and can in some cases be recommended for cytoskeletal antigens when the epitope is not readily accessible.
  • After PFA fixation, rinse with 50 mM ammonium chloride.

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.
Positive western blot control: anti-Keratin K5 antibody
Cat. No: 64003
  • Whole cell lysate from MCF7 human breast adenocarcinoma cell line
  • Validated and tested western blot control for: anti-Keratin K5 guinea pig polyclonal antibody, Cat. No. GP-CK5

$85.00*
anti-guinea pig IgG goat polyclonal, HRP conjugate
Cat. No: 90001
  • Goat polyclonal
  • Suitable for ELISA and WB
  • Reacts with guinea pig

$109.00*