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

Reviews (4)Q&A (3)References (20)
Western blot - Anti-PPAR alpha (phospho S12) antibody (ab3484)
  • Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)
  • Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)
  • Western blot - Anti-PPAR alpha (phospho S12) antibody (ab3484)
  • Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)
  • Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)

Key features and details

  • Rabbit polyclonal to PPAR alpha (phospho S12)
  • Suitable for: ICC, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

概述

  • 产品名称

    Anti-PPAR alpha (phospho S12)抗体
    参阅全部 PPAR alpha 一抗

  • 描述

    兔多克隆抗体to PPAR alpha (phospho S12)

  • 宿主

    Rabbit

  • 特异性

    The antibody is expected to bind both phospho and non phospho forms.

  • 经测试应用

    适用于: ICC, WBmore details

  • 种属反应性

    与反应: Mouse, Human
    预测可用于: Guinea pig, Dog

  • 免疫原

    Synthetic peptide corresponding to Mouse PPAR alpha aa 1-100 (phospho S12).

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 阳性对照

    • WB: human U-87, MCF7, MDA-MB-231, C2C12, HepG2, and mouse NIH-3T3 ICC: C2C12, 3T3-L1, U-87 MG cells

  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab3484于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC
1/100 - 1/500.
WB (2)
1/100 - 1/1000. Predicted molecular weight: 52 kDa.
说明
ICC
1/100 - 1/500.
WB
1/100 - 1/1000. Predicted molecular weight: 52 kDa.

靶标

  • 功能

    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.

  • 组织特异性

    Skeletal muscle, liver, heart and kidney.

  • 序列相似性

    Belongs to the nuclear hormone receptor family. NR1 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.

  • 细胞定位

    Nucleus.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • Western blot - Anti-PPAR alpha (phospho S12) antibody (ab3484)
    Western blot - Anti-PPAR alpha (phospho S12) antibody (ab3484)
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)
    Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)
    Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)

    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.

  • Western blot - Anti-PPAR alpha (phospho S12) antibody (ab3484)
    Western blot - Anti-PPAR alpha (phospho S12) antibody (ab3484)
    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
    All lanes : Goat anti-rabbit IgG (H+L) at 1/2500 dilution

    Predicted band size: 52 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)
    Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)
    Immunocytochemistry/ Immunofluorescence - Anti-PPAR alpha (phospho S12) antibody (ab3484)This image is courtesy of an anonymous Abreview
    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

文献 (20)

发表研究结果有使用 ab3484？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab3484 被引用在 20 文献中.

  • Li W  et al. Pemafibrate suppresses oxidative stress and apoptosis under cardiomyocyte ischemia-reperfusion injury in type 1 diabetes mellitus. Exp Ther Med 21:331 (2021). PubMed: 33732304
  • Hritzo B  et al. Late Health Effects of Partial Body Irradiation Injury in a Minipig Model Are Associated with Changes in Systemic and Cardiac IGF-1 Signaling. Int J Mol Sci 22:N/A (2021). PubMed: 33807089
  • Xie M  et al. Neddylation inhibitor MLN4924 has anti-HBV activity via modulating the ERK-HNF1a-C/EBPa-HNF4a axis. J Cell Mol Med 25:840-854 (2021). PubMed: 33263949
  • Tu J  et al. Gegen Qinlian Decoction Coordinately Regulates PPAR? and PPARa to Improve Glucose and Lipid Homeostasis in Diabetic Rats and Insulin Resistance 3T3-L1 Adipocytes. Front Pharmacol 11:811 (2020). PubMed: 32595495
  • Gámez-Belmonte R  et al. Deficiency in Tissue Non-Specific Alkaline Phosphatase Leads to Steatohepatitis in Mice Fed a High Fat Diet Similar to That Produced by a Methionine and Choline Deficient Diet. Int J Mol Sci 22:N/A (2020). PubMed: 33374541
View all Publications for this product

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1-7 of 7 Abreviews or Q&A

Western blot abreview for Anti-PPAR alpha (phospho S12) antibody

 Average
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (SHSY5Y)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
SHSY5Y
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More

Abcam user community

Verified customer

提交于 Mar 10 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PPAR alpha (phospho S12) antibody

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Hepatocellular carcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Vector lab H3300
Permeabilization
No
Specification
Hepatocellular carcinoma
Blocking step
0.5%BSA 1x casine as blocking agent for 5 minute(s) · Concentration: 0.5% · Temperature: RT°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

提交于 Sep 24 2020

Western blot abreview for Anti-PPAR alpha (phospho S12) antibody

 Good
Abreviews
abreview image
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
Read More

Dr. Chad Brocker

Verified customer

提交于 Apr 17 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-PPAR alpha (phospho S12) antibody

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Jul 14 2010

Question

Follow up to previous correspondance: Thank you for your suggestions. I was able to get a signal with all of the antibodies, but I am worried about the sizes of the bands. According to your data sheet, the size of ALL the bands is about 52 kD. However, only the PPARalpha (ab8934) detects a specific band of this size. The bands of the phosphorylated forms of PPARalpha (antibodies ab3484 and ab3485)are closer to 70 kD, but are quite specific, producing little background except during long exposures (1h). Are you completely sure that the sizes of the phosphorylated forms should be approx. 52 kD, or is it possible that they are larger, i.e. the 70 kD bands? I hope you are able to answer this last query, as I want to ascertain that the signal we are getting is the proper one. For you question concerning storage, the antibodies were aliquoted and stored in -20 C immediately upon arrival.

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Answer

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 like to send you replacements for these two antibodies as they should recognise well the 52kDa band. Can you please provide me with the order number used to purchase the antibodies and I will arrange replacements to be sent to you ASAP. I'm very sorry for the delay, thank you for your patience. I look forward to hearing from you soon,

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Question

Thank you for your mail. With the anti-PPARalpha (ab3484) we did not see any signal. With anti-PPARalpha (ab8934) we saw various background bands after 1 h exposure, but not one of the correct size. We loaded 60 micrograms of protein/lane. We did not try increasing the concentration of the primary antibodies, as 1:500 is already very high. Hope this is of some help. Thank you for your quick reply. The antibodies which did not function were: ab3484-100 (lot:52477) and ab8934-100 (lot: 44507). I used the antibodies in western blotting, diluted at 1:500 in 5%milk-0.05%Tween-PBS. Secondary antibodies were anti-rabbit(IgG)-HRP (Chemicon, has worked constantly in our lab)at 1:5000 dilution in 5%milk-0.05%Tween-PBS. The samples were mouse primary hepatocyte cell lysates (lysis buffer: 150 mM NaCl, 50 mM HEPES, 5mM EDTA, 0.1% NP-40 + protease/phosphatase inhibitors)which to my knowledge should contain large amounts of PPARalpha. Due to this fact, here were no positive controls (and we had no clue what would be a better positive control then hepatocyte lysates). I hope this is of some help to clear up the issue as these antibodies are critical in our studies.

Read More
Answer

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 the primary and incubating overnight at 4C. If you still do not see a signal, I will send you a replacement vial free of charge. For both, how were the antibodies stored after you received them?

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Question

Thank you for your quick reply. The antibodies which did not function were: ab3484-100 (lot:52477) and ab8934-100 (lot: 44507). I used the antibodies in western blotting, diluted at 1:500 in 5%milk-0.05%Tween-PBS. Secondary antibodies were anti-rabbit(IgG)-HRP (Chemicon, has worked constantly in our lab)at 1:5000 dilution in 5%milk-0.05%Tween-PBS. The samples were mouse primary hepatocyte cell lysates (lysis buffer: 150 mM NaCl, 50 mM HEPES, 5mM EDTA, 0.1% NP-40 + protease/phosphatase inhibitors)which to my knowledge should contain large amounts of PPARalpha. Due to this fact, here were no positive controls (and we had no clue what would be a better positive control then hepatocyte lysates). I hope this is of some help to clear up the issue as these antibodies are critical in our studies.

Read More
Answer

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 load? Did you try increasing the concentration of the primary antibodies? Thank you, and I look forward to hearing from you.

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