In-Cell ELISA
Validated antibodies, dyes and kits for
the analysis of fixed adherent and
suspension cells.
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In-Cell ELISA
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Advantages of In-Cell ELISA
Flexible
•
Measure specific protein level or post-translation modifications (e.g.
phosphorylation or cleavage events).
•
Suitable for adherent and non-adherent cell types.
•
Microplate format allows (1) parallel testing of low numbers of analytes across
a range of culture conditions, drug treatments, or cell lines (2) testing of many
analytes across few conditions.
•
Colorimetric or IR-dye detection methods (IR-dyes can be duplexed within wells).
Fast and inexpensive
•
No lysate preparation or lysate transfer required.
•
No capture antibody means less reagent cost than sandwich ELISAs.
•
96- and 384-well formats are high-throughput and primed for automation.
Reproducible, quantitative
•
•
•
Direct fixation in microplates freezes and preserves cells in their biologically
relevant state.
Suited for duplicate or triplicate measurements.
Signals can be normalized to total protein readout or to cell number.
Specific
•
•
•
MitoSciences’ antibodies are rigorously validated for ICE application.
Optimized assay kits available.
Support packs and a spectrum of detection antibodies simplify experimental
design.
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Summary of the method
In-Cell ELISA is a quantitative immunocytochemistry method to
measure protein levels or post-translational modifications of cells.
Key Features
•
•
•
•
•
•
Cells are cultured to approximately 80% confluency in a 96- or 384-well plate,
a drug or other treatment is applied to stimulate a cellular response.
Cells are fixed and permeabilized in the wells. Fixation to data acquisition
requires ~30 minutes of hands-on time. Alternatively, fixed cell plates are stable
for weeks to months at 4°C in the presence of sodium azide.
Cells are blocked and exposed to primary antibody which binds to their
intended targets within the mitochondria or other subcellular compartment. After
incubation, the unbound primary antibodies are washed away.
Secondary antibodies are added and the plates are scanned. Unbound
secondary antibody is washed away, reaction buffer is added for the
colorimetric assays and the signal is read on a suitable instrument for the kit
type.
MitoSciences anti-mouse and anti-rabbit HRP-conjugated secondary
antibodies, and also anti-mouse and anti-rabbit IRdye® conjugated secondary
antibodies are available for these assays.
Antibody signal can be normalized to Janus Green whole cell stain to account
for any differences in seeding density between wells.
ICE data can be collected with a LiCor® Odyssey® or Aerius® scanner using IRdye®labeled secondary antibodies or a standard absorbance microplate reader using
HRP-labeled secondary antibodies. When a LiCor® infrared reader is available,
duplexing readouts within a single well is possible using IR-800 and IR-680 labeled
secondary antibodies.
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Product Lists
ICE Kits
ICE (In-Cell ELISA) Support Pack
ICE (In-Cell ELISA) Support Pack w/o plates
MitoBiogenesis™ In-Cell ELISA Kit (IR)
MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric)
PARP-1 (cleaved) In-Cell ELISA Kit (IR)
PhosphoPDH In-Cell ELISA Kit (Colorimetric)
PhosphoPDH In-Cell ELISA Kit (IR)
ICE Validated antibodies
ACAA1
ACAA1
ACADM
ACADS
Aconitase 2
Aconitase 2
AGXT
AIF
ALDH2
ATPG
Catalase
Cleaved PARP
CPT2
CRYZ
Cyclophilin 40
Cytochrome C
DCXR
DECR1
ECH1
Epoxide hydrolase
FH
Frataxin
GAPDH
GOT2
HADHA
HADHB
Reacts with
Cow,
Cow,
Hu
Cow,
Cow,
Hu, Ms, Rat
Hu, Ms, Rat
Hu, Ms, Rat
Hu, Ms, Rat
Reacts with
Hu,
Hu
Hu,
Hu,
Hu,
Hu,
Hu,
Hu
Hu,
Hu,
Hu,
Hu
Hu,
Hu
Hu,
Hu,
Hu
Hu
Hu,
Hu
Hu,
Hu,
Hu
Hu,
Hu
Hu
Ms, Rat
Ms, Rat, Cow
Rat
Ms, Rat
Ms, Rat
Rat, Cow
Ms, Rat, Cow
Ms, Rat, Cow
Rat, Cow
Ms, Rat, Cow
Ms, Rat, Cow
Ms, Rat, Ce, Cow
Rat
Rat, Cow
Ms, Rat, Cow, Dog
Ms, Rat, Cow
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Amounts
5
5
2
2
2
2
2
x
x
x
x
x
x
x
96
96
96
96
96
96
96
tests
tests
tests
tests
tests
tests
tests
Product code
ab111542
ab111541
ab110216
ab110217
ab110215
ab110219
ab110218
Product code
ab110289
ab110290
ab110296
ab110318
ab110320
ab110321
ab110313
ab110327
ab110311
ab119686
ab110292
ab110315
ab110293
ab110310
ab110324
ab110325
ab110283
ab110287
ab110294
ab110307
ab110286
ab113691
ab110305
ab113693
ab110281
ab110301
ICE Validated antibodies (continued)
HADHSC
HMGCL
HSDL2
Hsp60
MADD
MDH2
MGST3
Mitochondrial Pyruvate dehydrogenase kinase 1
Mitofilin
mtTFA
Nitrotyrosine
OGDH
PCB
Pyruvate Dehydrogenase E1-alpha subunit
Pyruvate Dehydrogenase E2
SIRT1
SLIRP
Superoxide Dismutase 2
VCP
Reacts with
Hu
Hu
Hu
Hu
Hu,
Hu,
Hu,
Hu,
Hu
Hu
Hu,
Hu,
Hu,
Hu,
Hu,
Hu,
Hu
Hu,
Hu,
Ms, Rat
Ms, Rat, Pig
Rat
Cow
Ms, Rat, Cow
Cow, Pig
Ms, Rat, Cow
Ms, Rat, Cow
Cow
Ms, Rat
Ms, Rat, Cow
Ms, Rat
Product code
ab110284
ab110295
ab110298
ab110312
ab110316
ab110317
ab110309
ab110335
ab110329
ab119684
ab110282
ab110306
ab110314
ab110330
ab110332
ab110304
ab119687
ab110300
ab110308
IRdye® secondary antibodies and IRDye® kits offer greater dynamic range and
sensitivity. In addition, MitoSciences offers isotype-specific anti-mouse IRdye®
conjugated secondary antibodies to allow duplexing with two mouse monoclonal
antibodies of differing isotype. All secondary antibodies are included in ICE reagent
kits or can be purchased separately.
Goat
Goat
Goat
Goat
Goat
Goat
Goat
polyclonal
polyclonal
polyclonal
polyclonal
polyclonal
polyclonal
polyclonal
Secondary Antibody
Secondary Antibody
Secondary Antibody
Secondary Antibody
Secondary Antibody
Secondary Antibody
Secondary Antibody
Product code
to
to
to
to
to
to
to
IgG - H&L (IRDye® 800CW)
Rabbit IgG - H&L (IRDye® 680)
Mouse IgG1 (IRDye® 680LT)
IgG2a (IRDye® 800CW)
Mouse (IRDye® 800CW)
Mouse IgG - H&L (HRP)
Rabbit IgG - H&L (HRP)
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Copyright © 2012 Abcam, All Rights Reserved. The Abcam logo is a registered trademark.
All information / detail is correct at time of going to print.
ab110403
ab110404
ab110405
ab110406
ab110407
ab110408
ab110409
282_11_MBS
Secondary antibodies