Anti-FANCA/FAA 抗体 (ab5063)

Reviews (3)Q&A (2)References (9)

Key features and details

  • Rabbit polyclonal to FANCA/FAA
  • Suitable for: WB
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-FANCA/FAA antibody
    FANCA/FAA 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to FANCA/FAA

  • 由来種

    Rabbit

  • 特異性

    Detects a band at 130kDa in HeLa cell lysate that we believe corresponds to FANCA/FAA.

  • アプリケーション

    適用あり: WBmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide corresponding to Human FANCA/FAA aa 995-1009 conjugated to keyhole limpet haemocyanin.
    Sequence:

    SRSYDHSENSDLVFG-C


    Database link: O15360

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

  • 特記事項

     This product was previously labelled as FANCA

     

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

  • バッファー

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
  • 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

アプリケーション

Our Abpromise guarantee covers the use of ab5063 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
WB 1/500 - 1/2000. Detects a band of approximately 130 kDa (predicted molecular weight: 163 kDa).

ターゲット情報

  • 機能

    DNA repair protein that may operate in a postreplication repair or a cell cycle checkpoint function. May be involved in interstrand DNA cross-link repair and in the maintenance of normal chromosome stability.

  • 関連疾患

    Defects in FANCA are a cause of Fanconi anemia (FA) [MIM:227650]. FA is a genetically heterogeneous, autosomal recessive disorder characterized by progressive pancytopenia, a diverse assortment of congenital malformations, and a predisposition to the development of malignancies. At the cellular level it is associated with hypersensitivity to DNA-damaging agents, chromosomal instability (increased chromosome breakage), and defective DNA repair.

  • 翻訳後修飾

    Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylation is required for the formation of the nuclear complex. Not phosphorylated in cells derived from groups A, B, C, E, F, G, and H.

  • 細胞内局在

    Nucleus. Cytoplasm. The major form is nuclear. The minor form is cytoplasmic.
  • Information by UniProt

  • 参照データベース

    • 別名

      • FA 1 antibody
      • FA antibody
      • FA H antibody
      • FA1 antibody
      • FAA antibody
      • FACA antibody
      • FAH antibody
      • Fanca antibody
      • FANCA_HUMAN antibody
      • FANCH antibody
      • Fanconi anemia complementation group A antibody
      • Fanconi anemia complementation group H antibody
      • Fanconi anemia group A protein antibody
      • Fanconi anemia type 1 antibody
      • MGC75158 antibody
      • Protein FACA antibody
      see all

    画像

    • Western blot - Anti-FANCA/FAA antibody (ab5063)
      Western blot - Anti-FANCA/FAA antibody (ab5063)
      Anti-FANCA/FAA antibody (ab5063) at 1/1500 dilution + HeLa whole cell lysate at 35 µg

      Secondary
      IRDye 800 conjugated Goat anti-Rabbit IgG [H&L] at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size: 163 kDa
      Observed band size: 130 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 42 kDa, 75 kDa. We are unsure as to the identity of these extra bands.

    参考文献 (9)

    ab5063 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。

    ab5063 は 9 報の論文で使用されています。

    • Abdul-Sater Z  et al. FANCA safeguards interphase and mitosis during hematopoiesis in vivo. Exp Hematol 43:1031-1046.e12 (2015). IF . PubMed: 26366677
    • Vanuytsel K  et al. FANCA knockout in human embryonic stem cells causes a severe growth disadvantage. Stem Cell Res 13:240-50 (2014). WB . PubMed: 25108529
    • Rio P  et al. Targeted gene therapy and cell reprogramming in Fanconi anemia. EMBO Mol Med 6:835-48 (2014). WB ; Human . PubMed: 24859981
    • Shen C  et al. Regulation of FANCD2 by the mTOR pathway contributes to the resistance of cancer cells to DNA double-strand breaks. Cancer Res 73:3393-401 (2013). PubMed: 23633493
    • Meier D & Schindler D Fanconi anemia core complex gene promoters harbor conserved transcription regulatory elements. PLoS One 6:e22911 (2011). WB . PubMed: 21826217
    • Lu WT  et al. The Fanconi anemia pathway is downregulated upon macrophage differentiation through two distinct mechanisms. Cell Cycle 10: (2011). WB ; Human . PubMed: 21926477
    • Tulpule A  et al. Knockdown of Fanconi anemia genes in human embryonic stem cells reveals early developmental defects in the hematopoietic lineage. Blood 115:3453-62 (2010). WB ; Human . PubMed: 20089964
    • Mattera L  et al. The E1A-associated p400 protein modulates cell fate decisions by the regulation of ROS homeostasis. PLoS Genet 6:e1000983 (2010). WB ; Human . PubMed: 20548951
    • Wilson JB  et al. FANCG promotes formation of a newly identified protein complex containing BRCA2, FANCD2 and XRCC3. Oncogene 27:3641-52 (2008). PubMed: 18212739

    レビューと Q&A

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    1-5 of 5 Abreviews or Q&A

    Question

    Anbei der vollständig ausgefüllte Fragebogen!



    Vielen dank!

    Mit besten grüßen

    1) Abcam product code ab 96089, ab 5063, ab54676, ab54645, ab 6264

    2) Abcam order reference number or product batch number

    3) Description of the problem: see pictures below, either to high cross reactivity or no signal detectable

    4) Sample preparation:
    Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysate
    Lysis buffer: RIPA
    Protease inhibitors: sigma (http://www.sigmaaldrich.com/catalog/product/sigma/p8340?lang=de®ion=AT)
    Phosphatase inhibitors: sigma
    Reducing agent
    Boiling for ≥5 min? yes/no yes
    Protein loaded ug/lane or cells/lane 15-100 µg
    Positive control yes
    Negative control

    5) Percentage of gel 10%
    Type of membrane Nylon
    Protein transfer verified yes (Ponseau red)
    Blocking agent and concentration 5% BSA in 1xTBS/T (as well as 5% milk in 1xTBS/T but in milk no signal could be detected)
    Blocking time 4 °C over night, as well as 1 h room temperature
    Blocking temperature see above

    6) Primary antibody (If more than one was used, describe in “additional notes”) :
    Concentration or dilution: FancG 1:500, FancA 1:1000, XPD 1:1000, CSB 1:1500
    Diluent buffer 5% BSA as well as milk in 1xTBS/T
    Incubation time over night
    Incubation temperature: 4°C

    7) Secondary antibody:
    Species:
    Reacts against:
    Concentration or dilution 1:1000
    Diluent buffer 5% BSA in 1x TBS/T
    Incubation time: 1h
    Incubation temperature: room temperature
    Fluorochrome or enzyme conjugate: HRP

    8) Washing after primary and secondary antibodies:
    Buffer 1x TBS/T
    Number of washes 3

    9)Detection method
    10) How many times have you run this staining?
    Do you obtain the same results every time?
    What steps have you altered to try and optimize the use of this antibody?
    * changed incubation time
    * changes milk to BSA
    * blocked over night or 1h

    Read More
    Answer

    Vielen Dank für Ihren Anruf und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

    Es tut mir leid, dass Sie Probleme mit unseren Antikörper hatten.

    Um Ihr Problem zu lösen, möchte ich die folgenden Vorschläge zu Veränderung Ihres Protokolls machen, bzw noch ein paar zusätzliche Fragen stellen:

    1. Um was für Zelllinien handelt es sich hier, bzw aus welcher Spezies wurden die Zellen ursprünglich gewonnen? Sind die Zellen transfektiert?

    2. Was ist Ihre Positiv- Kontrolle?

    3.Verwenden Sie einen Anti-Mouse Sekundär- AKim falle der ab54645 und ab54676?

    4. Verwenden Sie immer einen frischen Blot pro Antikörper oder strippen Sie diesen zwischendurch?

    5. Haben Sie Ihre Zellen transfektiert oder wollen Sie das endogene Protein detektieren? Für den ersten Fall senken Sie den Proteingehalt auf 5- 10ug pro Bahn ab, im zweiten Fall gehen Sie runter auf 20 ug.

    6. Filtern Sie die Blockmittelllösung, dadurch sollten die schwarzen Punkte verschwinden und die Ergebnisse klarer werden.

    7. Senken Sie die Konzentration der Sekundär- Antikörper auf mindestens 1/5000 ab.

    8. Blockieren Sie weiterhin eine Stunde und inkubieren sie in der höchsten von uns empfohlenenVerdünnung über Nacht bei 4C.



    Bitte lassen Sie mich wissen, ob ich Ihnen helfen konnte und zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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    Question

    long list of abs, two orders, only two work!
    works either not at all or strong cross-reactivity

    Read More
    Answer

    Vielen Dank für Ihren Anruf.

    Es tut mir leid zu hören, dass Sie Probleme mit diesen Antikörpern haben.

    Ich habe unseren Fragebogen als Word-Dokument an diese E-Mail angehängt. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und möglichst Veränderungsvorschläge machen, die Ihre Ergebnisse verbessern werden. Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und er innerhalb der letzten 180 Tage gekauft wurde, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.


    Ich freue mich, bald wieder von Ihnen zu hören.

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    Western blot abreview for Anti-FANCA/FAA antibody

     Below Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    50 µg
    Specification
    HeLa
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Aug 14 2007

    Western blot abreview for Anti-FANCA/FAA antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (THP1 - Human acute monocytic leukemia cell)
    Loading amount
    150 µg
    Specification
    THP1 - Human acute monocytic leukemia cell
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
    Read More

    Dr. Wei-Ting Lu

    Verified customer

    投稿 Jul 24 2007

    Western blot abreview for Anti-FANCA/FAA antibody

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Cal51 & SW620)
    Loading amount
    750 cells
    Specification
    Cal51 & SW620
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
    Read More

    Mr. Richard Allum

    Verified customer

    投稿 Oct 24 2006

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