Product datasheet
Anti-PPAR alpha antibody [3B6/PPAR] - ChIP Grade
ab2779
2 Abreviews 12 References 6 Images
Overview
Product name
Anti-PPAR alpha antibody [3B6/PPAR] - ChIP Grade
Description
Mouse monoclonal [3B6/PPAR] to PPAR alpha - ChIP Grade
Tested applications
ChIP, Flow Cyt, IP, WB, IHC-Fr
Species reactivity
Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Human
Immunogen
Recombinant full length protein corresponding to PPAR alpha.
Positive control
Skeletal muscle, heart, liver, kidney, or mouse adipose tissue extract. 3T3 cells, Jurkat cells,
HeLa cells.
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 long
term. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.05% Sodium azide
Constituent: 99% PBS
Purity
IgG fraction
Clonality
Monoclonal
Clone number
3B6/PPAR
Isotype
IgG2b
Applications
Our Abpromise guarantee covers the use of ab2779 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
ChIP
Use at an assay dependent concentration. PubMed: 20599735
Flow Cyt
Use 1µg for 106 cells.
IP
Use at an assay dependent concentration. PubMed: 16051671
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Application
Abreviews
WB
Notes
Use a concentration of 2 - 5 µg/ml. Detects a band of approximately 52 kDa
(predicted molecular weight: 52 kDa).
In some cases it might necessary to perform an immunoprecipitation prior to the
WB (see Sumanasekera et al).
In case no band observed check the IgG2b specificity of secondary antibody.
IHC-Fr
1/400.
Target
Function
Ligand-activated transcription factor. Key regulator of lipid metabolism. Activated by the
endogenous ligand 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine (16:0/18:1-GPC).
Activated by oleylethanolamide, a naturally occurring lipid that regulates satiety (By similarity).
Receptor for peroxisome proliferators such as hypolipidemic drugs and fatty acids. Regulates
the peroxisomal beta-oxidation pathway of fatty acids. Functions as transcription activator for the
ACOX1 and P450 genes. Transactivation activity requires heterodimerization with RXRA and is
antagonized by NR2C2.
Tissue specificity
Skeletal muscle, liver, heart and kidney.
Sequence similarities
Belongs to the nuclear hormone receptor family. NR1 subfamily.
Contains 1 nuclear receptor DNA-binding domain.
Cellular localization
Nucleus.
Anti-PPAR alpha antibody [3B6/PPAR] - ChIP Grade images
ChIP assay with HepG2 cells via real time
PCR calculation. Levels of binding of PPARα
with the human C3 promoter, apoA-I promoter
(positive control), and NOD1 gene (negative
control) are shown for chromatin precipitates
with antibodies against PPARα (ab2779) or
β-actin (ab3280). Values are presented as
means ± S.E. (error bars) of four independent
experiments. The statistical analyses of
ChIP - Anti-PPAR alpha [3B6/PPAR] antibody -
differences between compared groups were
ChIP Grade (ab2779)
performed using an unpaired Student's t test
Image from Mogilenko DA et al., J Biol Chem.
2013;288(3):1726-38. Fig 2(D).; doi:
10.1074/jbc.M112.437525.
(*, p < 0.05).
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Lanes 1 - 2 : Anti-PPAR alpha antibody
[3B6/PPAR] - ChIP Grade (ab2779) at
1/1000 dilution
Lane 3 : Anti-PPAR alpha antibody
[3B6/PPAR] - ChIP Grade (ab2779) at 1/500
dilution
Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : NIH-3T3 cell lysate
Western blot - Anti-PPAR alpha antibody
Lysates/proteins at 25 µg per lane.
[3B6/PPAR] - ChIP Grade (ab2779)
Predicted band size : 52 kDa
Observed band size : 52 kDa
Flow cytometry analysis of PPAR alpha
showing positive staining in the nucleus of
Jurkat cells (ab7899) compared to an isotype
control (blue). Cells were harvested, adjusted
to a concentration of 1-5x10^6 cells/ml, fixed
with 2% paraformaldehyde, washed with
PBS, and incubated with ab2779 (0.25
ug/test) for 60 min at room temperature. Cells
were then blocked in a solution of 2% BSAPBS for 30 min at room temperature,
incubated for 40 min at room temperature in
Flow Cytometry - Anti-PPAR alpha antibody
the dark using a Dylight 488-conjugated goat
[3B6/PPAR] - ChIP Grade (ab2779)
anti-mouse IgG (H+L) secondary antibody
(ab96879), and re-suspended in PBS for
FACS analysis.
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Flow cytometry analysis of PPAR alpha
showing positive staining in the nucleus of
HeLa cells (ab150035) compared to an
isotype control (blue). Cells were harvested,
adjusted to a concentration of 1-5x10^6
cells/ml, fixed with 2% paraformaldehyde,
washed with PBS, and incubated
with ab2779 (0.25 ug/test) for 60 min at room
temperature. Cells were then blocked in a
solution of 2% BSA-PBS for 30 min at room
temperature, incubated for 40 min at room
Flow Cytometry - Anti-PPAR alpha antibody
temperature in the dark using a Dylight 488-
[3B6/PPAR] - ChIP Grade (ab2779)
conjugated goat anti-mouse IgG (H+L)
secondary antibody (ab96879), and resuspended in PBS for FACS analysis.
Flow cytometry analysis of PPAR alpha
showing positive staining in the nucleus of
3T3 cells (ab7179) compared to an isotype
control (blue). Cells were harvested, adjusted
to a concentration of 1-5x10^6 cells/ml, fixed
with 2% paraformaldehyde, washed with
PBS, and incubated with ab2779 (0.25
ug/test) for 60 min at room temperature. Cells
were then blocked in a solution of 2% BSAPBS for 30 min at room temperature,
incubated for 40 min at room temperature in
Flow Cytometry - Anti-PPAR alpha antibody
the dark using a Dylight 488-conjugated goat
[3B6/PPAR] - ChIP Grade (ab2779)
anti-mouse IgG (H+L) secondary antibody
(ab96879), and re-suspended in PBS for
FACS analysis.
4
Overlay histogram showing HepG2 cells
stained with ab2779 (red line). The cells were
fixed with 4% paraformaldehyde (10 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. The cells were then incubated
Flow Cytometry-Anti-PPAR alpha antibody
[3B6/PPAR] - ChIP Grade(ab2779)
with the antibody (ab2779, 1µg/1x106 cells)
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 IgG2b [PLPV219] (ab91366,
2µg/1x106 cells ) used under the same
conditions. Acquisition of >5,000 events was
performed. This antibody gave a positive
signal in HepG2 cells fixed with 80%
methanol (5 min) /permeabilized in 0.1%
PBS-Tween used under the same conditions.
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