The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50 - 1/75. An incubation period of 30 minutes at room temperature is recommended. Formalin fixed paraffin embedded tissue sections require high temperature antigen unmasking with 10 mM citrate buffer, pH 6.0 prior to immunostaining.
Use at an assay dependent concentration. PMID: 17065488
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Involvement in disease
Defects in KRT5 are a cause of epidermolysis bullosa simplex Dowling-Meara type (DM-EBS) [MIM:131760]. DM-EBS is a severe form of intraepidermal epidermolysis bullosa characterized by generalized herpetiform blistering, milia formation, dystrophic nails, and mucous membrane involvement. Defects in KRT5 are the cause of epidermolysis bullosa simplex with migratory circinate erythema (EBSMCE) [MIM:609352]. EBSMCE is a form of intraepidermal epidermolysis bullosa characterized by unusual migratory circinate erythema. Skin lesions appear from birth primarily on the hands, feet, and legs but spare nails, ocular epithelia and mucosae. Lesions heal with brown pigmentation but no scarring. Electron microscopy findings are distinct from those seen in the DM-EBS, with no evidence of tonofilament clumping. Defects in KRT5 are a cause of epidermolysis bullosa simplex Weber-Cockayne type (WC-EBS) [MIM:131800]. WC-EBS is a form of intraepidermal epidermolysis bullosa characterized by blistering limited to palmar and plantar areas of the skin. Defects in KRT5 are a cause of epidermolysis bullosa simplex Koebner type (K-EBS) [MIM:131900]. K-EBS is a form of intraepidermal epidermolysis bullosa characterized by generalized skin blistering. The phenotype is not fundamentally distinct from the Dowling-Meara type, althought it is less severe. Defects in KRT5 are the cause of epidermolysis bullosa simplex with mottled pigmentation (MP-EBS) [MIM:131960]. MP-EBS is a form of intraepidermal epidermolysis bullosa characterized by blistering at acral sites and 'mottled' pigmentation of the trunk and proximal extremities with hyper- and hypopigmentation macules. Defects in KRT5 are the cause of Dowling-Degos disease (DDD) [MIM:179850]; also known as Dowling-Degos-Kitamura disease or reticulate acropigmentation of Kitamura. DDD is an autosomal dominant genodermatosis. Affected individuals develop a postpubertal reticulate hyperpigmentation that is progressive and disfiguring, and small hyperkeratotic dark brown papules that affect mainly the flexures and great skin folds. Patients usually show no abnormalities of the hair or nails.
Immunocytochemistry - Anti-Cytokeratin 5 antibody [XM26] (ab17130)This image is courtesy of an anonymous Abreview
ab17130 staining Cytokeratin 5 in Human keratinocytes by ICC (Immunocytochemistry). Cells were fixed with ethanol:acetone (1:1), permeabilized with Triton X 100 (0.1% for 5 minutes) and blocked with FCS/H2O2 3% (endo. PO) for 5 minutes at 21°C. Samples were incubated with primary antibody (1/100 in FBS Stain-Buffer) for 1 hour at 21°C. An undiluted HRP-conjugated Goat anti-rabbit/mouse IgG polyclonal was used as the secondary antibody. Negative cells were stained with Mayer's hematoxilin.
Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 5 antibody [XM26] (ab17130)Image courtesy of an anonymous Abreview.
ab171330 staining Cytokeratin 5 in human airway epithelial cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in formaldehyde, permeabilized using PBS/ 0.25% Triton for 10 minutes, blocked with 5% horse serum for 1 hour at room temperature and then incubated with ab17130 at a 1/200 dilution for 15 hours. The secondary used was an Alexa-Fluor 488 conjugated goat anti-mouse polyclonal used at a 1/750 dilution.
Overlay histogram showing A431 cells stained with ab17130 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab17130, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Butler CR et al. Vacuum-assisted decellularization: an accelerated protocol to generate tissue-engineered human tracheal scaffolds. Biomaterials124:95-105 (2017).
IHC-P, IHC - Wholemount
Read more (PubMed: 28189871) »
Crowley C et al. Surface modification of a POSS-nanocomposite material to enhance cellular integration of a synthetic bioscaffold. Biomaterials83:283-93 (2016).
Read more (PubMed: 26790147) »