Product datasheet
Anti-pan Keratin antibody [80] ab8068
4 Abreviews 11 References 5 Images
Overview
Product name
Anti-pan Keratin antibody [80]
Description
Mouse monoclonal [80] to pan Keratin
Specificity
In immunoblots most human keratins are recognized. No reactivity with neurofilament, vimentin
GFAP or Desmin
Tested applications
Flow Cyt, IHC-Fr, IHC-P, WB, ICC/IF
Species reactivity
Reacts with: Mouse, Rat, Human
Immunogen
Callus cytokeratins isolated from fresh human skin tissue
Positive control
Human callus or keratinocytes
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.07% Sodium azide
Constituent: 1% Fetal calf serum
Purity
Tissue culture supernatant
Clonality
Monoclonal
Clone number
80
Isotype
IgG1
Applications
Our Abpromise guarantee covers the use of ab8068 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Flow Cyt
Abreviews
Notes
1/20. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control
with this antibody.
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Application
Abreviews
IHC-Fr
Notes
Use at an assay dependent concentration. Stains all types of keratin containing
cells (epithelia) in frozen sections of various tissues, with the exception of
myoepithelial cells.
IHC-P
Use at an assay dependent concentration. For paraffin embedded tissue a TUF
pretreatment is recommended.
WB
Use at an assay dependent concentration. Detects a band of approximately 65
kDa (predicted molecular weight: 65 kDa).
ICC/IF
Use a concentration of 1 µg/ml.
Target
Relevance
Cytokeratin is a family of basic and acidic proteins, present in dermal tissues. Each cytokeratin
is formed by heterotetramers of different types of keratin, that span from 1 to 18. In keratinized
epidermis, 50 kD keratin is present in the basal layer, while 56.5 kD keratin is present in
suprabasal layers, where as 58 kD keratin is present in the basal and suprabasal layers, while
65 to 67 kD keratin is present in the cells above the basal layers.
Cellular localization
Cytoplasmic
Anti-pan Keratin antibody [80] images
ab8068 staining human foreskin tissue
sections by Immunohistochemistry
(Formalin/PFA-fixed paraffin embedded).
Tissue underwent fixation in
paraformaldehyde, heat mediated antigen
retrieval in 10mM Citrate buffer pH 6.0 in
boiling water bath for 10 minutes, followed by
cooling at room temperature for 30 minutes.
The blocking was done using 1%BSA/5%
Immunohistochemistry (Formalin/PFA-fixed
normal donkey serum in PBS pH 7.4 for 2
paraffin-embedded sections) - pan Keratin
hours at room temperature. The primary
antibody [80] (ab8068)
antibody, diluted 1/10 (PBS pH 7.4, 1%BSA,
This image is courtesy of an anonymous Abreview
0.1% sodium azide) and incubated with
sample for 16 hours at 4°C. A HRPconjugated donkey polyclonal to mouse Ig,
diluted 1/1000 was used as the secondary.
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ICC/IF image of ab8068 stained HeLa 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% PBSTween for 1h to permeabilise the cells and
block non-specific protein-protein
interactions. The cells were then incubated
with the antibody (ab8068, 1µg/ml) overnight
at +4°C. The secondary antibody (green) was
Alexa Fluor® 488 goat anti-mouse IgG (H+L)
used at a 1/1000 dilution for 1h. Alexa Fluor®
Immunocytochemistry/ Immunofluorescence -
594 WGA was used to label plasma
Anti-pan Keratin antibody [80] (ab8068)
membranes (red) at a 1/200 dilution for 1h.
DAPI was used to stain the cell nuclei (blue)
at a concentration of 1.43µM.
Anti-pan Keratin antibody [80] (ab8068) at
1/250 dilution + A431 (Human epithelial
carcinoma cell line) Whole Cell Lysate at 10
µg
Secondary
Goat Anti-Mouse IgG H&L (HRP)
preadsorbed (ab97040) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Western blot - pan Keratin antibody [80] (ab8068)
Predicted band size : 65 kDa
Observed band size : 65 kDa
Additional bands at : 49 kDa,58 kDa. We
are unsure as to the identity of these extra
bands.
Exposure time : 30 seconds
3
Overlay histogram showing A431 cells
stained with ab8068 (red line). The cells were
fixed with 80% methanol (5 min) and then
permeabilized with 0.1% PBS-Triton 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 (ab8068, 1/20
Flow Cytometry - Anti-pan Keratin antibody [80]
dilution) for 30 min at 22°C. The secondary
(ab8068)
antibody used was DyLight® 488 goat antimouse IgG (H+L) (ab96879) at 1/500 dilution
for 30 min at 22°C. 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.
ab8068 staining pan Keratin in rat
glioblastoma cell line C6 by
Immunocytochemistry/ Immunofluorescence.
Cells were fixed in paraformaldehyde,
permeabilized using 0,1% Triton X 100 in
PBS, blocked with 0.5% BSA for 30 minutes
Immunocytochemistry/ Immunofluorescence -
at room temperature and then incubated with
Anti-pan Keratin antibody [80] (ab8068)
ab8068 at a 1/50 dilution for 16 hours at 4°C.
Image courtesy of an anonymous Abreview.
The secondary used was a Cy3 conjugated
goat anti-mouse polyclonal used at a 1/400
dilution. Nuclei are counterstained with DAPI.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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