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Anti-CYP7A1 抗体 (ab65596)

Reviews (4)Q&A (2)References (54)
Western blot - Anti-CYP7A1 antibody (ab65596)

    Key features and details

    • Rabbit polyclonal to CYP7A1
    • Suitable for: WB, ELISA
    • Reacts with: Mouse
    • Isotype: IgG

    製品の概要

    • 製品名

      Anti-CYP7A1 antibody
      CYP7A1 一次抗体 製品一覧

    • 製品の詳細

      Rabbit polyclonal to CYP7A1

    • 由来種

      Rabbit

    • 特異性

      From May 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help.

    • アプリケーション

      適用あり: WB, ELISAmore details

    • 種交差性

      交差種: Mouse
      交差が予測される動物種: Rat, Dog, Human, Pig, Rhesus monkey

    • 免疫原

      Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human CYP7A1.

      Immunogen の所有権に関して (Peptide available as ab69726.)

    • ポジティブ･コントロール

      • WB: High Fat Diet and regular Mouse Liver lysate

    • 特記事項

      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

    製品の特性

    アプリケーション

    The Abpromise guarantee

    Abpromise保証は、 次のテスト済みアプリケーションにおけるab65596の使用に適用されます

    アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

    アプリケーション Abreviews 特記事項
    WB (4)
    Use a concentration of 1 µg/ml. Predicted molecular weight: 57 kDa.
    ELISA
    Use at an assay dependent concentration.
    特記事項
    WB
    Use a concentration of 1 µg/ml. Predicted molecular weight: 57 kDa.
    ELISA
    Use at an assay dependent concentration.

    ターゲット情報

    • 機能

      Catalyzes a rate-limiting step in cholesterol catabolism and bile acid biosynthesis by introducing a hydrophilic moiety at position 7 of cholesterol. Important for cholesterol homeostasis.

    • 組織特異性

      Detected in liver.

    • パスウェイ

      Lipid metabolism; bile acid biosynthesis.

    • 配列類似性

      Belongs to the cytochrome P450 family.

    • 細胞内局在

      Endoplasmic reticulum membrane. Microsome membrane.
    • Information by UniProt

    • 参照データベース

      see all

    • 別名

      • Cholesterol 7 alpha hydroxylase antibody
      • Cholesterol 7 alpha monooxygenase antibody
      • Cholesterol 7-alpha-hydroxylase antibody
      • Cholesterol 7-alpha-monooxygenase antibody
      • CP7A antibody
      • CP7A1_HUMAN antibody
      • CYP 7 antibody
      • CYP7 antibody
      • CYP7A1 antibody
      • CYPVII antibody
      • Cytochrome P450 7A1 antibody
      • Cytochrome P450, family 7, subfamily A, polypeptide 1 antibody
      • Cytochrome P450, subfamily VIIA (cholesterol 7 alpha monooxygenase), antibody
      • Cytochrome P450, subfamily VIIA (cholesterol 7 alpha-monooxygenase), polypeptide 1 antibody
      • MGC126826 antibody
      • MGC138389 antibody
      see all

    画像

    • Western blot - Anti-CYP7A1 antibody (ab65596)
      Western blot - Anti-CYP7A1 antibody (ab65596)
      All lanes : Anti-CYP7A1 antibody (ab65596) at 1 µg/ml

      Lanes 1 & 5 : High Fat Diet Mouse Liver lysate
      Lanes 2 & 6 : Mouse Liver lysate
      Lanes 3 & 7 : Mouse Thymus lysate
      Lanes 4 & 8 : Mouse Colon lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 57 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Primary antibody (Green) - ab65596 without (Lanes 1-4) and with (Lanes 5-8) immunising peptide. 

      Loading Control (Red) - ab8245 at 0.1µg/ml, all lanes.

      Secondary antibody - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed at 1/10000 dilution and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed at 1/10000 Dilution.

      Blocking Buffer - 3% Milk

    参考文献 (54)

    ab65596 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。

    ab65596 は 54 報の論文で使用されています。

    • Liu F  et al. Nigakinone alleviates DSS-induced experimental colitis via regulating bile acid profile and FXR/NLRP3 signaling pathways. Phytother Res 37:15-34 (2023). PubMed: 36054406
    • Xu W  et al. L-Theanine regulates lipid metabolism by modulating gut microbiota and bile acid metabolism. J Sci Food Agric 103:1283-1293 (2023). PubMed: 36087337
    • Chen MY  et al. Psoralen induces liver injury and affects hepatic bile acids metabolism in female and male C57BL/6J mice. Phytother Res 37:2280-2289 (2023). PubMed: 36724888
    • Li H  et al. Geniposide plus chlorogenic acid reverses non-alcoholic steatohepatitis via regulation of gut microbiota and bile acid signaling in a mouse model in vivo. Front Pharmacol 14:1148737 (2023). PubMed: 37077819
    • Lin X  et al. Intermittent Fasting Alleviates Non-Alcoholic Steatohepatitis by Regulating Bile Acid Metabolism and Promoting Fecal Bile Acid Excretion in High-Fat and High-Cholesterol Diet Fed Mice. Mol Nutr Food Res 67:e2200595 (2023). PubMed: 37148502
    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

    Western blot abreview for Anti-CYP7A1 antibody

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Liver)
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Loading amount
    50 µg
    Specification
    Liver
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Apr 22 2020

    Western blot abreview for Anti-CYP7A1 antibody

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (liver)
    Gel Running Conditions
    Non-reduced Denaturing (12%)
    Loading amount
    30 µg
    Specification
    liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Jul 05 2017

    Western blot abreview for Anti-CYP7A1 antibody

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (7.5)
    Sample
    Rat Tissue lysate - whole (liver)
    Specification
    liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7.5%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Sep 09 2013

    Western blot abreview for Anti-CYP7A1 antibody

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (liver)
    Loading amount
    30 µg
    Specification
    liver
    Gel Running Conditions
    Reduced Denaturing (4-20% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Jan 19 2011

    Question

    Quería consultar si se ha testado en western blot de tejido de rata el anticuerpo ab65596 (anti-CYP7A1) y si lo poseen en stock.

    Read More
    Answer



    ab65596 es un llamado “Fast Track” antibody. Se trata de anticuerpos que no se han caracterizado completamente, y que se han añadido al catálogo antes de realizar la validación completa por la necesidad urgente de añadir anticuerpos contra la proteína concreta.

    Gracias a que no están completamente caracterizados su precio es reducido y no están cubiertos por la garantía Abpromise. Solo se han testado en el laboratorio en ELISA, y cualquier publicación o referencia de los usuarios del mismo se compensa con Abpoints extra canjeables.

    Los términos y condiciones de estos anticuerpos los podéis consultar en su datasheet.

    Por tanto, en respuesta a tu pregunta, este anticuerpo no se ha usado en Western Blot, ni con tejidos de ratón, y tampoco podemos ofrecer códigos de descuento Abtrial. Sin embargo, si estáis interesados en usarlo en esta especie / aplicación, y enviarnos el Abreview, sí que a cambio os beneficiareis de 950 Abpoints en vuestra cuenta. El anticuerpo está actualmente en stock.

    En cualquier caso, hay disponibles otros dos anticuerpos que si se han validado en WB con lisados de ratón: ab106060 y ab65596.

    Read More

    Question

    Dear Technical support,

    I hope this email finds you well.

    We've received a product complaint which we are hoping that you will be able to help address. Attached is the completed questionnaire.

    With many thanks and best regards,



    1) Abcam product code: ab65596



    2) Abcam order reference number or product batch number: 831470



    3) Description of the problem: There seems to be bands in the beginning. After that, the WB film either turns black entirely or no band shows up.



    4) Sample preparation

    Type of sample: whole cell lysates

    Lysis buffer: RIPA

    Protease inhibitors: Halt(tm)Protease & PhosphataseInhibitor Cocktail

    Phosphatase inhibitors: Halt(tm)Protease & PhosphataseInhibitor Cocktail

    Reducing agent: Loading buffer (Bio-Rad) & β-mercaptoethanol

    Boiling for ≥5 min? Yes, 95˚C for 15 minutes

    Protein loaded: 50ug / lane

    Positive control: HepG2, rat liver

    Negative control: Blank lane



    5) Percentage of gel: 8% separating. 5% stacking.

    Type of membrane: PVDF

    Protein transfer verified : Yes, using ladder as a gauge

    Blocking agent and concentration: 5% milk in 1X TBS w/ 0.05% Tween 20

    Blocking time: 1 hour

    Blocking temperature: Room temperature



    6) Primary antibody

    Concentration or dilution: 1:1000, 1:1500, 1:2000

    Diluent buffer: 1X TBS w/ 0.05% Tween 20

    Incubation time: Overnight

    Incubation temperature: 4˚C




    7) Secondary antibody

    Species: Goat

    Reacts against: anti-rabbit

    Concentration or dilution: 1:10000, 1:15000, 1:20000

    Diluent buffer: 1X TBS w/ 0.05% Tween 20

    Incubation time: 1 hour

    Incubation temperature: Room temperature

    Fluorochrome or enzyme conjugate: Enzyme conjugate



    8) Washing after primary and secondary antibodies:

    Buffer: twice with 1X TBS w/ 0.05% Tween 20, thrice with 1X TBS

    Number of washes: 5, 5 minutes each wash



    9) Detection method: ECL



    10) How many times have you run this staining? Seven

    Do you obtain the same results every time? No, in the beginning got bands, after that, WB film either turns black entirely or no bands at all.

    What steps have you altered to try and optimize the use of this antibody?


    Primary antibody

    Secondary antibody

    Blocking


    Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

    I have attached film picture when there were bands (or so I think, that they should be the correct bands). I did not attach those that are entirely black or entirely blank (w/o any bands).

    Read More
    Answer

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

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    Reviewing this case, I would like to offer some suggestions to help optimize the results from ab65596. I would also appreciate if you can confirm some further details:

    1. I am sorry to confirm that this is a FastTrack antibody, not yet tested and guaranteed for rat or western blot. As our FastTrack antibodies have not been fully characterized at Abcam, we cannot guarantee that the antibody will work in any application other than ELISA against the immunizing peptide and I am sorry they are therefore not covered by the full Abpromise guarantee. Therefore, we are regrettably not able to provide a credit or refund on this occasion.

    As an alternative, the customer may like to consider submitting an Abreview, for which they can obtain 900 Abpoints for data ona Fast Track antibody.

    2. The secondary antibody mentioned on the form is stated as an anti-mouse. Could you confirm if this is correct? Please provide the supplier and catalog code for the secondary antibody.

    As ab65596 is a rabbit polyclonal you will need to use an anti-rabbit IgG secondary antibody. An anti-mouse secondary will not work for detection of the rabbit polyclonal primary.

    3. Is the secondary antibody working well with other primary antibodies? If you are getting background from a correct secondary, I can suggest it would be important to include a no primary control to assess if the background is from non specific binding of the secondary. The concentration of secondary may need to be reduced in order to optimize the results.

    I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More

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