Overview

  • Product name
    Anti-alpha 1 Fetoprotein antibody [AFP-01]
    See all alpha 1 Fetoprotein primary antibodies
  • Description
    Mouse monoclonal [AFP-01] to alpha 1 Fetoprotein
  • Host species
    Mouse
  • Specificity
    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.
  • Tested applications
    Suitable 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

  • Function
    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.
  • Tissue specificity
    Plasma. Synthesized by the fetal liver and yolk sac.
  • Sequence similarities
    Belongs to the ALB/AFP/VDB family.
    Contains 3 albumin domains.
  • Developmental stage
    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.
  • 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 localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

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
    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

References

This product has been referenced in:
  • Shafa M  et al. Human-Induced Pluripotent Stem Cells Manufactured Using a Current Good Manufacturing Practice-Compliant Process Differentiate Into Clinically Relevant Cells From Three Germ Layers. Front Med (Lausanne) 5:69 (2018). Read more (PubMed: 29600249) »
  • Naumann M  et al. Impaired DNA damage response signaling by FUS-NLS mutations leads to neurodegeneration and FUS aggregate formation. Nat Commun 9:335 (2018). ICC/IF . Read more (PubMed: 29362359) »
See all 17 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

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

Orla Fannon

Verified customer

Submitted Sep 12 2017

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

Abcam user community

Verified customer

Submitted Aug 31 2015

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
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

Abcam user community

Verified customer

Submitted Feb 06 2013

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

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

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

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

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

Sign up