Anti-PPAR alpha (phospho S12) antibody (ab3484)

Overview

  • Product name
    Anti-PPAR alpha (phospho S12) antibody
    See all PPAR alpha primary antibodies
  • Description
    Rabbit polyclonal to PPAR alpha (phospho S12)
  • Specificity
    The antibody is expected to bind both phospho and non phospho forms.
  • Tested applications
    Suitable for: ICC/IF, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Guinea pig, Dog
  • Immunogen

    Synthetic peptide corresponding to Mouse PPAR alpha aa 8-19.
    Sequence: ICPLSpPLEADDL
    (Peptide available as ab4998)

Applications

Our Abpromise guarantee covers the use of ab3484 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
EMSA Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB 1/100 - 1/1000. Predicted molecular weight: 52 kDa.Can be blocked with PPAR alpha peptide (ab4998).
Flow Cyt Use 3-5µg for 106 cells.

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.
  • 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

Images

  • Immunofluorescent analysis of Phospho-PPAR alpha pSer12 (green) showing staining in the nucleus of C2C12 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Phospho-PPAR alpha pSer12 polyclonal antibody (ab3484) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • All lanes : Anti-PPAR alpha (phospho S12) antibody (ab3484) at 1/200 dilution

    Lane 1 : C2C12 cell lysate
    Lane 2 : NIH-3T3 cell lysate
    Lane 3 : 3T3-L1 cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 25 µg per lane.


    Predicted band size : 52 kDa
    Observed band size : 52 kDa
  • Immunofluorescent analysis of Phospho-PPAR alpha pSer12 (green) showing staining in the nucleus of 3T3-L1 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Phospho-PPAR alpha pSer12 polyclonal antibody (ab3484) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • All lanes : Anti-PPAR alpha (phospho S12) antibody (ab3484) at 1/1000 dilution

    Lane 1 : U-87 MG with Skimmed milk
    Lane 2 : MCF7 with Skimmed milk
    Lane 3 : MDA-MB-231 with Skimmed milk
    Lane 4 : C2C12 with Skimmed milk
    Lane 5 : Hep G2 with Skimmed milk
    Lane 6 : NIH/3T3 with Skimmed milk

    Lysates/proteins at 20 µg per lane.
    Blocking peptides at 5 % per lane.

    Secondary
    Goat anti-rabbit IgG (H+L) at 1/2500 dilution

    Predicted band size : 52 kDa
  • Immunofluorescent analysis of Phospho-PPAR alpha pSer12 (green) showing staining in the nucleus of U-87 MG cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Phospho-PPAR alpha pSer12 polyclonal antibody (ab3484) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • ab3484 staining PPAR alpha (phospho S12) in Mouse neuronal cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/100 in PBS) for 18 hours at 4°C. A Cy2®-conjugated Donkey anti-rabbit IgG polyclonal (1/100) was used as the secondary antibody.

    See Abreview

  • ab3484 staining PPAR alpha (phospho S12) in MCF7 cells by Flow Cytometry. The sample was incubated with the primary antibody (3-5 ug/million cells in 2.5% BSA) for 2 hours at room temperature. An Alexa Fluor® 488-conjugated Goat anti-rabbit was used as the secondary antibody (1/400). Red histogram represents ab3484, pink histogram represents isotype control, purple histogram represents unstained control and green histogram represents no primary antibody control.

References

This product has been referenced in:
  • Azimzadeh O  et al. A dose-dependent perturbation in cardiac energy metabolism is linked to radiation-induced ischemic heart disease in Mayak nuclear workers. Oncotarget 8:9067-9078 (2017). WB . Read more (PubMed: 27391067) »
  • Chen Z & Wang Q Activation of PPAR? by baicalin attenuates pulmonary hypertension in an infant rat model by suppressing HMGB1/RAGE signaling. FEBS Open Bio 7:477-484 (2017). Read more (PubMed: 28396833) »

See all 6 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Mouse Tissue lysate - nuclear (Liver nuclei lysates from Ppara null and wildtype)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
Liver nuclei lysates from Ppara null and wildtype
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Dr. Chad Brocker

Verified customer

Submitted Apr 17 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (neuronal cells)
Specification
neuronal cells
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 14 2010

Thank you for your recent e-mail. My colleague Jennifer is unexpectedly away this week and I would like to help you. I understand you have recurring problems with ab3484 and ab3485 detecting the wrong bands and your protocol seems fine. I would lik...

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Thanks again for your email. At this point I would like to make the following suggestions that will hopefully help you out. For ab8934 I suggest using 3T3 whole cell lysate as a positive control. For ab3484, I suggest increasing the concentration of th...

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Thank you for your email and the details in which you have provided. Can you please tell me - what exactly are you seeing on your blots? Are you not getting any signal at all or are you seeing bands but not at the correct size? How much protein did you...

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