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

Overview

  • Product nameAnti-alpha 1 Fetoprotein antibody [AFP-01]
    See all alpha 1 Fetoprotein primary antibodies
  • Description
    Mouse monoclonal [AFP-01] to alpha 1 Fetoprotein
  • SpecificityThe 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.
  • Tested applicationsSuitable for: Sandwich ELISA, ICC/IF, ELISA, IP, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

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

  • Positive control
    • 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.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • FunctionBinds 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.
  • Tissue specificityPlasma. Synthesized by the fetal liver and yolk sac.
  • Sequence similaritiesBelongs to the ALB/AFP/VDB family.
    Contains 3 albumin domains.
  • Developmental stageOccurs 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.
  • Post-translational
    modifications
    Independent studies suggest heterogeneity of the N-terminal sequence of the mature protein and of the cleavage site of the signal sequence.
    Sulfated.
  • Cellular localizationSecreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • Afp 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

Anti-alpha 1 Fetoprotein antibody [AFP-01] images

  • 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.

  • 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
    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

    This image is courtesy of an anonymous abreview.

    See Abreview

References for Anti-alpha 1 Fetoprotein antibody [AFP-01] (ab3980)

This product has been referenced in:
  • Nie Y  et al. Scalable passaging of adherent human pluripotent stem cells. PLoS One 9:e88012 (2014). ICC/IF ; Human . Read more (PubMed: 24498239) »
  • Pettinato G  et al. ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells. PLoS One 9:e100742 (2014). ICC/IF ; Human . Read more (PubMed: 25365581) »

See all 7 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

Submitted Aug 31 2015

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...

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Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Feb 06 2013

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...

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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...

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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 fu...

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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 recomme...

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