製品の概要

  • 製品名Anti-SPHK1 antibody
    SPHK1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to SPHK1
  • アプリケーション適用あり: ICC/IF, IHC-P, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Non Human Primates
  • 免疫原

    Synthetic peptide corresponding to Human SPHK1 aa 286-315 (internal sequence) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: 8877
    (Peptide available as ab187382)

  • ポジティブ・コントロール
    • WB: 293, HepG2, HUVEC, Raji, C6 and C2C12 cell lysates. Mouse kidney tissue lysate. IHC-P: Human hepatocarcinoma tissue. ICC/IF: HeLa cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab71700 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 a concentration of 5 µg/ml.
IHC-P 1/50 - 1/100.
WB 1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 43 kDa).Can be blocked with SPHK1 peptide (ab187382).

There are two known isoforms of this protein, one predicted around 43 kDa and the other around 51 kDa.

ターゲット情報

Anti-SPHK1 antibody 画像

  • All lanes : Anti-SPHK1 antibody (ab71700) at 1/1000 dilution

    Lane 1 : 293 cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : HUVEC cell lysate
    Lane 4 : Raji cell lysate
    Lane 5 : C6 cell lysate
    Lane 6 : C2C12 cell lysate

    Lysates/proteins at 35 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG (H+L) at 1/5000 dilution

    Predicted band size : 43 kDa
  • Anti-SPHK1 antibody (ab71700) at 1/500 dilution + Lysate prepared from human MDA MB 231 cells at 15 µg

    Secondary
    HRP-conjugated donkey polyclonal to rabbit IgG at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 43 kDa
    Observed band size : 48,52 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes

    This image is a courtesy of Anonymous Abreview.

    See Abreview

  • Anti-SPHK1 antibody (ab71700) at 1/1000 dilution + mouse kidney tissue lysate at 12.5 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/5000 dilution

    Predicted band size : 43 kDa
    Observed band size : 48 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 52 kDa. We are unsure as to the identity of these extra bands.

    Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical staining of SPHK1 in Human hepatocarcinoma tissue sections (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections) with ab71700 at a dilution of 1/25. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hours at 38°C. Antigen retrieval was heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A HRP-conjugated goat anti-rabbit polyclonal (ready to use) was used as the secondary antibody.

  • ICC/IF image of ab71700 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab71700, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Anti-SPHK1 antibody (ab71700) 使用論文

This product has been referenced in:
  • Testai FD  et al. Changes in the metabolism of sphingolipids after subarachnoid hemorrhage. J Neurosci Res 93:796-805 (2015). WB ; Human . Read more (PubMed: 25597763) »
  • Cai K  et al. Silencing diacylglycerol kinase-theta expression reduces steroid hormone biosynthesis and cholesterol metabolism in human adrenocortical cells. Biochim Biophys Acta 1841:552-62 (2014). Human . Read more (PubMed: 24369117) »

See all 5 Publications for this product

Product Wall

We have previously been in contact regarding ab71700, which was not providing satisfactory results.

A free of charge replacement vial was issued and I am writing to inquire if the replacement product has worked. I would appreciate if you cou...

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Merci de nous avoir contactés.

Nous sommes désolés d'apprendre que le produit que vous avez reçu ne fonctionne pas comme attendu. Le numéro de commande de remplacement gratuit avec la référenc...

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Merci d'avoir pris le temps de remplir notre questionnaire et de nous avoir contactés.
Comme nous avons discuté au téléphone, ces informations sont très importantes pour nous. Ils nous permettent d'abord de cible...

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Merci pour votre appel ce matin. Je suis désolée d'apprendre aussi que ab71700 ne vous donne pas de bons résultats en immunohistochimie.

Veuillez trouver ci-joint lequestionnaire discuté au téléphone ...

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According to our records, ab71700 was proving difficult to use inWB and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand ...

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I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that w...

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Thanks for your email. Sorry to hear this vial of ab71700 is giving you trouble.
If you have purchased this antibody within the last 6 months or so I would be happy to provide a free replacement antibody. In your reply, please include your PO numb...

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Thank you for providing this further information.

I appreciate your cooperation and understand your concerns. It is regrettable the results have not been succesful.

In order to help with our invesigation of this case before decidi...

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Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this...

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Thank you for contacting us and reporting the problems observed with lot inconsistency with Anti-SPHK1 antibody (ab71700).


I am currently looking into whether there has been any lot inconsistency observed with this antibody and if we po...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"