Product datasheet
Anti-HMW Cytokeratin antibody [34betaE12] ab86725
1 Abreviews 5 References 3 Images
Overview
Product name
Anti-HMW Cytokeratin antibody [34betaE12]
Description
Mouse monoclonal [34betaE12] to HMW Cytokeratin
Specificity
ab86725 recognizes cytokeratin 1, 5, 10 and 14
Tested applications
IHC-P, ICC/IF, Flow Cyt
Species reactivity
Reacts with: Mouse, Rat, Dog, Human
Immunogen
HMW CK 34betaE12
Positive control
Human, mouse or rat skin, prostate, squamous cell carcinoma or basal cell carcinoma tissue.
This antibody gave a positive result when used in the following methanol fixed cell lines: DU145.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze /
thaw cycle.
Storage buffer
Preservative: 0.09% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.2-7.4
Clonality
Monoclonal
Clone number
34betaE12
Isotype
IgG1
Light chain type
kappa
Applications
Our Abpromise guarantee covers the use of ab86725 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Notes
IHC-P
1/50 - 1/100.
ICC/IF
Use a concentration of 10 µg/ml.
Flow Cyt
1/100. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype
control with this antibody.
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Target
Relevance
Cytokeratins are intermediate filament keratins found in the intracytoplasmic cytoskeleton of
epithelial tissue. There are two types of Cytokeratins: the low weight, acidic type I cytokeratins
and the high weight, basic or neutral type II. Cytokeratins are usually found in pairs comprising a
type I Cytokeratin and a type II cytokeratin. The high molecular weight cytokeratins, which are the
basic or neutral cytokeratins, comprise subtypes CK1 (67), CK2 (65.5), CK3 (64), CK4 (59),
CK5 (58), CK6 (56), CK7 (54), CK8 (52.5) and CK9. The low molecular weight cytokeratins,
which are the acidic cytokeratins, comprise subtypes CK10 (56.5), CK12 (56), CK13 (53),
CK14 (50), CK16( 48), CK17 (46), CK18 (45), CK19(48) and CK20(46).
Cellular localization
Cytoplasmic
Anti-HMW Cytokeratin antibody [34betaE12] images
ab86725, at 1/50 dilution, staining Human
HMW Cytokeratin in prostate, using
Immunohistochemistry, Formalin/PFA-fixed
paraffin embedded tissue.
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) - HMW Cytokeratin
antibody [34betaE12] (ab86725)
Overlay histogram showing A431 cells
stained with ab86725 (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
Flow Cytometry-Anti-HMW Cytokeratin antibody
[34betaE12](ab86725)
(ab86725, 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.
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ICC/IF image of ab86725 stained DU145
cells. The cells were 100% methanol fixed (5
min) and then incubated in 1%BSA / 10%
normal goat serum / 0.3M glycine in 0.1%
PBS-Tween for 1h to permeabilise the cells
and block non-specific protein-protein
interactions. The cells were then incubated
with the antibody ab86725 at 10µg/ml
overnight at +4°C. The secondary antibody
(green) was DyLight® 488 goat anti- mouse
(ab96879) IgG (H+L) used at a 1/250 dilution
Immunocytochemistry/ Immunofluorescence -
for 1h. Alexa Fluor® 594 WGA was used to
Anti-HMW Cytokeratin antibody [34betaE12]
label plasma membranes (red) at a 1/200
(ab86725)
dilution for 1h. DAPI was used to stain the cell
nuclei (blue) at a concentration of 1.43µM.
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