Anti-alpha 1 Fetoprotein抗体[AFP-01] (ab3980)

Reviews (3)Q&A (5)References (29)

Key features and details

  • Mouse monoclonal [AFP-01] to alpha 1 Fetoprotein
  • Suitable for: Sandwich ELISA, ICC/IF, ELISA, IP, WB
  • Reacts with: Human
  • Isotype: IgG1

概述

  • 产品名称

    Anti-alpha 1 Fetoprotein抗体[AFP-01]
    参阅全部 alpha 1 Fetoprotein 一抗

  • 描述

    小鼠单克隆抗体[AFP-01] to alpha 1 Fetoprotein

  • 宿主

    Mouse

  • 特异性

    The antibody reacts with human alpha-Fetoprotein (AFP), an oncofetal antigen of 70 kDa. AFP is a major fetal plasma protein, but is not present in healthy adult tissues. Elevated AFP concentrations in adult plasma may be an early marker of hepatocellular carcinoma or teratoblastoma, while high concentrations in amniotic fluid may indicate severe congenital defects of a fetus.

  • 经测试应用

    适用于: Sandwich ELISA, ICC/IF, ELISA, IP, WBmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Full length native protein (purified) corresponding to Human alpha 1 Fetoprotein.

  • 阳性对照

    • Hep G2 human hepatocellular carcinoma cell line Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with non-reducing/reducing Laemmli SDS-PAGE sample buffer.

性能

应用

Our Abpromise guarantee covers the use of ab3980 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Sandwich ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration. Use at an assay dependant dilution. Interaction of the antibody with AFP is dependent on the presence of calcium ions (strongly inhibited by chelating agents). Such characteristics of the antibody can be exploited for immunoaffinity purification of APF under mild elution conditions.
WB Use at an assay dependent concentration. Detects a band of approximately 70 kDa.

靶标

  • 功能

    Binds copper, nickel, and fatty acids as well as, and bilirubin less well than, serum albumin. Only a small percentage (less than 2%) of the human AFP shows estrogen-binding properties.

  • 组织特异性

    Plasma. Synthesized by the fetal liver and yolk sac.

  • 序列相似性

    Belongs to the ALB/AFP/VDB family.
    Contains 3 albumin domains.

  • 发展阶段

    Occurs in the plasma of fetuses more than 4 weeks old, reaches the highest levels during the 12th-16th week of gestation, and drops to trace amounts after birth. The serum level in adults is usually less than 40 ng/ml. AFP occurs also at high levels in the plasma and ascitic fluid of adults with hepatoma.

  • 翻译后修饰

    Independent studies suggest heterogeneity of the N-terminal sequence of the mature protein and of the cleavage site of the signal sequence.
    Sulfated.

  • 细胞定位

    Secreted.
  • Information by UniProt

  • 数据库链接

    • 别名

      • Afp antibody
      • AFPD antibody
      • Alpha fetoglobulin antibody
      • Alpha fetoprotein antibody
      • Alpha fetoprotein precursor antibody
      • Alpha-1-fetoprotein antibody
      • Alpha-fetoglobulin antibody
      • Alpha-fetoprotein antibody
      • alpha-fetoprotein, Hereditary persistence of, included antibody
      • FETA antibody
      • FETA_HUMAN antibody
      • Hereditary persistence of alpha fetoprotein antibody
      • HPAFP antibody
      see all

    图片

    • Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Fetoprotein antibody [AFP-01] (ab3980)
      Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Fetoprotein antibody [AFP-01] (ab3980)

      ICC/IF image of ab3980 stained MCF7 cells. The cells were 100% methanol fixed (5 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 (ab3980, 5µg/ml) overnight at +4°C. The secondary antibody (green)ÿwas Alexa Fluor® 488 goat anti-mouse 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.

    • Western blot - Anti-alpha 1 Fetoprotein antibody [AFP-01] (ab3980)
      Western blot - Anti-alpha 1 Fetoprotein antibody [AFP-01] (ab3980)This image is courtesy of an anonymous abreview.
      All lanes : Anti-alpha 1 Fetoprotein antibody [AFP-01] (ab3980) at 1/1000 dilution

      Lane 1 : Lanes 1 human embryonic stem (ES) cell lysates.
      Lane 2 : Lanes 2 human embryonic stem (ES) cell lysates.
      Lane 3 : Lanes 3 definitive endoderm lysates.
      Lane 4 : Lanes 4 definitive endoderm lysates.
      Lane 5 : Lanes 5 hepatoblast lysates.
      Lane 6 : Lanes 6 hepatoblast lysates.
      Lane 7 : Lanes 7 hepatocyte lysates.
      Lane 8 : Lanes 8 hepatocyte lysates.

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-mouse HRP (Dako) at 1/1000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Observed band size: 62-65 kDa
      why is the actual band size different from the predicted?


      Exposure time: 15 minutes

      See Abreview

    文献 (29)

    发表研究结果有使用 ab3980？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab3980 被引用在 29 文献中.

    • Lenz D  et al. Generation of an iPSC line from a patient with infantile liver failure syndrome 2 due to mutations in NBAS: DHMCi004-A. Stem Cell Res 35:101398 (2019). PubMed: 30772683
    • Wei J  et al. Establishment and characterization of the GC-030-35 cell line derived from gastric hepatoid adenocarcinoma. Cancer Manag Res 11:1275-1287 (2019). PubMed: 30799954
    • Jung-Klawitter S  et al. Generation of 2 iPSC clones from a patient with DNAJC12 deficiency: DHMCi003-A and DHMCi003-B. Stem Cell Res 36:101402 (2019). PubMed: 30901742
    • Lenz D  et al. Generation of an induced pluripotent stem cell (iPSC) line, DHMCi005-A, from a patient with CALFAN syndrome due to mutations in SCYL1. Stem Cell Res 37:101428 (2019). PubMed: 30959346
    • Tongkobpetch S  et al. Generation of two human iPSC lines (MDCUi001-A and MDCUi001-B) from dermal fibroblasts of a Thai patient with X-linked osteogenesis imperfecta using integration-free Sendai virus. Stem Cell Res 39:101493 (2019). PubMed: 31326747
    View all Publications for this product

    客户评价及客户问答

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    1-8 of 8 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-alpha 1 Fetoprotein antibody [AFP-01]

     Inconclusive
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (mES derived embryoid bodies)
    Permeabilization
    Yes - 0.05% triton
    Specification
    mES derived embryoid bodies
    Blocking step
    Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 20% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Orla Fannon

    Verified customer

    提交于 Sep 12 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-alpha 1 Fetoprotein antibody [AFP-01]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HEK293)
    Permeabilization
    No
    Specification
    HEK293
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Acetone
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 31 2015

    Question

    I need a pair of anti alpha fetoprotein antibodies for ELISA. But I do not know which ones could work together. Can you give me some suggestion?

    Read More
    Answer

    Having reviewed our catalog of anti-alpha 1 Fetoprotein antibodies suitable for use in ELISA, I have found the following 2 pairs which have been verified by us. AB8276 pairs with ab37207. More information about these items may be found at: Ab8276: https://www.abcam.com/index.html?datasheet=8276 Ab37207: https://www.abcam.com/index.html?datasheet=37207 Another pair which have been verified to work with each other in ELISA are ab3980 and ab3969. More information about these products may be found at: Ab3980: https://www.abcam.com/index.html?datasheet=3980 Ab3969: https://www.abcam.com/index.html?datasheet=3969

    Read More

    Western blot abreview for Anti-alpha 1 Fetoprotein antibody [AFP-01]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (hES-derived hepatocytes)
    Loading amount
    10 µg
    Specification
    hES-derived hepatocytes
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    提交于 Feb 06 2013

    Question

    We have been using your goat anti-human AFP antibody and I noticed on your website that it is no longer sold in the US. Do you offer a substitute or have a recommendation on what to use instead of ab8202-1?

    Read More
    Answer

    I am sorry for the inconvenience caused by this import restriction. While ab3980 is our only Anti-alpha 1 Fetoprotein raised in goat, we do offer a number of other Anti-alpha 1 Fetoprotein. We would recommend use of ab3980, Mouse monoclonal [AFP-01] to alpha 1 Fetoprotein as an alternative to the restricted antibody. Ab3980 has been tested in many of the same applications and has the same immunogen. This product also has been referenced in four publications.

    Read More

    Question

    I am guessing I received a bad batch. In any case, we would like to use a different antibody to avoid further problems. I see that the Anti-alpha 1 Fetoprotein antibody [AFP-11] (ab3969) has been tested exactly in the application we need to use it. Could you send us this antibody? when could we expect to get it?

    Kind regards

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1098413.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    PS: The product will be dispatched tomorrow for Friday delivery.

    Read More

    Question

    yIn a rush and the secondary antibody isAlexa Fluor 568 donkey anti - mouseIgG (invitrogen A10037) not the onei mentioned before. I used a red secondary
    I knowalexa 488 is green andalexa 568 is red andi used the right filter, let's just not keep on going on this point please. The filteri used was correct .
    Looking forward to you reply
    Best Wishes

    Read More
    Answer

    ers used and secondary antibody are correct. We always double check the protocol steps with our customer when some of them do not make sense. I apologize for any inconvenience.

    This antibody has been tested in ICC/IF with MCF7 cells and is fully guaranteed; the product should have worked however we are unsure about the problem. In order to help you further I am happy providing free of charge replacement product or a different antibody. Could you let me know how you would like to proceed.

    Looking forward to hearing from you soon.

    Read More

    Question

    a h Anti-alpha 1 Fetoprotein antibody (ab3980).
    Details as follow:
    LOT NUMBER: GR39372-3
    ORDER NUMBER: 304583
    DESCRIPTION OF THE PROBLEM
    No staining or very low stain
    SAMPLE
    Human hepatocytes derivated from iPSC
    Huh7 cell ines for positive control
    PRIMARY ANTIBODY Anti-alpha 1 Fetoprotein antibody [AFP-01] (ab3980)
    diluent: blocking buffer (0.5% BSA in PBS); dilution: tested at 1/50, 1/100 and 1/200; Incubation time: o/n at 4º; wash step: TBS 3x5 minuts each
    DETECTION METHOD
    Fluorescence microscope
    POSITIVE AND NEGATIVE CONTROLS USED
    positive: Huh7 Hepatocyte cell line (image attached)
    negative: Fibroblasts
    ANTIBODY STORAGE CONDITIONS
    shipped at 4º, aliquot and stored at -20º
    FIXATION OF SAMPLE
    PFA 4% for 15 min
    ANTIGEN RETRIEVAL
    Cell were immediately stained after fixation, no antigen retrieval was needed
    PERMEABILIZATION STEP
    0.1% Triton X-100 PBS
    BLOCKING CONDITIONS
    1h at RT in 0.5% BSA in PBS
    SECONDARY ANTIBODY
    tested with:
    Alexa 488 Donkey anti-mouse (invitrogen)
    diluent: blocking buffer (0.5% BSA in PBS)
    dilution: 1/1000
    Incubation time: 1 hour at RT
    wash step: PBS 3x5 minuts each
    HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3
    HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes
    DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes
    Pictures enclosed (dilution 1 in 50)

    Read More
    Answer

    ram very sorry to hear that you are experiencing problems with this antibody.

    The protocol looks fine to me however I have following further suggestions

    - We used methanol as fixative when we characterized this antibody. We recommend trying this.
    - It is mentioned in the protocol that Alexa 488 conjugated secondary was used, Alexa 488 is green in colour however the images shows red staining which could mean that wrong filters were used for observing the images. Could you try using correct filters.

    I hope these suggestions will be helpful. Should you have any problem using this antibody again, please do not hesitate to contact me.

    Read More

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