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  1. Link

    ppar-alpha-phospho-s12-antibody-ab3484.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families Zinc Finger
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Anti-PPAR alpha (phospho S12) 抗体 (ab3484)

  • Datasheet
Reviews (3)Q&A (3)References (15)

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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)
  • Flow Cytometry - Anti-PPAR alpha (phospho S12) antibody (ab3484)

Key features and details

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

こちらの製品もご検討ください

二次抗体
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
タンパク質
Recombinant Human PPAR alpha protein (ab81927)

関連製品

製品の概要

  • 製品名

    Anti-PPAR alpha (phospho S12) antibody
    PPAR alpha 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to PPAR alpha (phospho S12)
  • 由来種

    Rabbit
  • 特異性

    The antibody is expected to bind both phospho and non phospho forms.
  • アプリケーション

    適用あり: ICC/IF, WB, Flow Cytmore details
  • 種交差性

    交差種: Mouse, Human
    交差が予測される動物種: Guinea pig, Dog
  • 免疫原

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

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 特記事項

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • 精製度

    Immunogen affinity purified
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Zinc Finger
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • Nuclear hormone receptors
    • Other
    • Cardiovascular
    • Lipids / Lipoproteins
    • Fatty Acids
    • Metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of lipids and lipoproteins
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial Biogenesis
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Nucleotide metabolism
    • Molecular processes
    • Mitochondrial transcription
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipid metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Fatty acids
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
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    • Metabolism
    • Types of disease
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    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Fatty acid oxidation

関連製品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human PPAR alpha protein (ab81927)

アプリケーション

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.

アプリケーション Abreviews 特記事項
ICC/IF Use at an assay dependent concentration.
WB 1/100 - 1/1000. Predicted molecular weight: 52 kDa.
Flow Cyt Use 3-5µg for 106 cells.

ターゲット情報

  • 機能

    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.
  • 組織特異性

    Skeletal muscle, liver, heart and kidney.
  • 配列類似性

    Belongs to the nuclear hormone receptor family. NR1 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • 細胞内局在

    Nucleus.
  • Target information above from: UniProt accession Q07869 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 403654 Dog
    • Entrez Gene: 100135604 Guinea pig
    • Entrez Gene: 5465 Human
    • Entrez Gene: 19013 Mouse
    • Omim: 170998 Human
    • SwissProt: Q95N78 Dog
    • SwissProt: O35507 Guinea pig
    • SwissProt: Q07869 Human
    • SwissProt: P23204 Mouse
    • Unigene: 103110 Human
    • Unigene: 710044 Human
    • Unigene: 212789 Mouse
    see all
  • 別名

    • 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

画像

  • 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

  • Flow Cytometry - Anti-PPAR alpha (phospho S12) antibody (ab3484)
    Flow Cytometry - Anti-PPAR alpha (phospho S12) antibody (ab3484)

    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.

プロトコール

  • Flow cytometry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

データシートおよび資料

    • Datasheet
  • 参考文献 (15)

    ab3484 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab3484 は 15 報の論文で使用されています。

    • Wang H  et al. Phytanic acid activates PPARa to promote beige adipogenic differentiation of preadipocytes. J Nutr Biochem 67:201-211 (2019). PubMed: 30951974
    • Sun YL  et al. Mechanism of pomegranate ellagic polyphenols reducing insulin resistance on gestational diabetes mellitus rats. Am J Transl Res 11:5487-5500 (2019). PubMed: 31632524
    • Barjaktarovic Z  et al. Hyperacetylation of Cardiac Mitochondrial Proteins Is Associated with Metabolic Impairment and Sirtuin Downregulation after Chronic Total Body Irradiation of ApoE -/- Mice. Int J Mol Sci 20:N/A (2019). PubMed: 31652604
    • Xiao Y  et al. Neurotensin contributes to pediatric intestinal failure-associated liver disease via regulating intestinal bile acids uptake. EBioMedicine 35:133-141 (2018). PubMed: 30104181
    • Yang BY  et al. Different physiological roles of insulin receptors in mediating nutrient metabolism in zebrafish. Am J Physiol Endocrinol Metab 315:E38-E51 (2018). PubMed: 29351486
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-6 of 6 Abreviews or Q&A

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

    Good
    Abreviews
    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

    投稿 Sep 24 2020

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

    Good
    Abreviews
    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

    投稿 Apr 17 2015

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

    Good
    Abreviews
    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

    投稿 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.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 09 2004

    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,

    Read More

    Abcam Scientific Support

    Answered on Nov 16 2004

    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

    Abcam community

    Verified customer

    Asked on Oct 29 2004

    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?

    Read More

    Abcam Scientific Support

    Answered on Nov 02 2004

    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

    Abcam community

    Verified customer

    Asked on Oct 25 2004

    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.

    Read More

    Abcam Scientific Support

    Answered on Oct 28 2004

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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