Anti-PPAR alpha antibody [3B6/PPAR] - ChIP Grade (ab2779)

Overview

  • Product nameAnti-PPAR alpha antibody [3B6/PPAR] - ChIP Grade
    See all PPAR alpha primary antibodies
  • Description
    Mouse monoclonal [3B6/PPAR] to PPAR alpha - ChIP Grade
  • Tested applicationsSuitable for: ChIP, Flow Cyt, IP, WB, IHC-Frmore details
  • 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.

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
WB 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

  • FunctionLigand-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 specificitySkeletal muscle, liver, heart and kidney.
  • Sequence similaritiesBelongs to the nuclear hormone receptor family. NR1 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • hPPAR antibody
    • MGC2237 antibody
    • MGC2452 antibody
    • NR1C1 antibody
    • Nuclear receptor subfamily 1 group C member 1 antibody
    • OTTHUMP00000197740 antibody
    • OTTHUMP00000197741 antibody
    • Peroxisome proliferative activated receptor alpha antibody
    • Peroxisome proliferator activated receptor alpha antibody
    • Peroxisome proliferator-activated receptor alpha antibody
    • PPAR antibody
    • PPAR-alpha antibody
    • ppara antibody
    • PPARA_HUMAN antibody
    • PPARalpha antibody
    see all

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 differences between compared groups were performed using an unpaired Student's t test (*, p < 0.05).

  • 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

    Lysates/proteins at 25 µg per lane.


    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% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody (ab96879), and re-suspended in PBS for FACS analysis.

  • 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 temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody (ab96879), and re-suspended 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% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody (ab96879), and re-suspended in PBS for FACS analysis.

  • 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 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.

References for Anti-PPAR alpha antibody [3B6/PPAR] - ChIP Grade (ab2779)

This product has been referenced in:
  • Abo Al Hayja M  et al. Reduced expression of peroxisome proliferator-activated receptor alpha in BAL and blood T cells of non-löfgren's sarcoidosis patients. J Inflamm (Lond) 12:28 (2015). Flow Cyt ; Human . Read more (PubMed: 25969669) »
  • Ambrosino P  et al. Functional and biochemical interaction between PPARa receptors and TRPV1 channels: Potential role in PPARa agonists-mediated analgesia. Pharmacol Res 87:113-22 (2014). Read more (PubMed: 25014183) »

See all 12 Publications for this product

Product Wall

Gracias por contactarnos de nuevo.

He llevado a cabo una búsqueda de anticuerpos que pudieran serviros para detectar las proteínas indicadas por WB en lisados de células procedentes de conejo.

Contra PPAR alpha ...

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Thank you for contacting us.
Regarding PPAR-delta ab:
I think ab58137 (www.abcam.com/ab58137) as you suggested would be the best antibody to try as it has been tested in flow cyt and using the same secondary ab (ab96879) would be fine. Howeve...

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Thank you for your enquiry. I am happy to help answer your questions.

1. I can confirm that ab2779 PPAR alpha antibody [3B6/PPAR] is tested and therefore covered by our 6 month guarantee for use in flow cytometry and human samples. So it sho...

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Thank you for your reply.

Given that ab2779 is a monoclonal antibody, we would expect there to be little batch to batch variability. It is sometimes observed in cases of contamination, but it is more likely that the epitope is detected on mul...

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Thank you for your reply and for resending your images. I agree that the background makes your results difficult to interpret. Given that the non-specific bands are of similar intensity to the band of interest, it is unlikely that protocol changes will...

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Thank you for contacting us. I am sorry this antibody is producing multiple bands in your experiment.

Unfortunately, I am unable to open the image that you attached. Could you please resend the image as a jpg or png file? Without seeing the i...

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Thanks for your enquiry.

We regularly ship antibodies on Saturday for Monday delivery. The products are quite stable and there should not be any loss of activity from this short time at elevated temperature. We have conducted in-house testing...

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I'm happy to help! Best of luck with your experiments.

As requested, I have issued a free of charge replacement for one vial of ab119416.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replaceme...

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Thanks for your reply.

I am happy to send you a vial of ab119416 as as replacement. I'm sorry if I forgot to ask you before but could you please forward the PO number or Abcam order reference number assiciated with the initial purchase of ab2...

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