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RecombinantRabMAb

Recombinant Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)

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Western blot - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
  • Flow Cytometry (Intracellular) - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
  • Immunoprecipitation - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
  • Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPAFP61] to alpha 1 Fetoprotein - Low endotoxin, Azide free
  • Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
  • Reacts with: Human

Overview

  • Product name

    Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free
    See all alpha 1 Fetoprotein primary antibodies

  • Description

    Rabbit monoclonal [EPAFP61] to alpha 1 Fetoprotein - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein corresponding to Human alpha 1 Fetoprotein.
    Database link: P02771

  • Positive control

    • Flow Cyt (intra): HepG2 cells; IP: HepG2 whole cell lysate; ICC/IF: HepG2 cells; IHC-P: Human liver cancer tissue; WB: HepG2 whole cell lysate.

  • General notes

    ab215379 is the carrier-free version of ab133617.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab215379 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
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376- Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Detects a band of approximately 70 kDa (predicted molecular weight: 68 kDa).
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Heat up to 98°C, below boiling, and then let cool for 10-20 min.
ICC/IF
Use at an assay dependent concentration.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376- Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Detects a band of approximately 70 kDa (predicted molecular weight: 68 kDa).
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Heat up to 98°C, below boiling, and then let cool for 10-20 min.
ICC/IF
Use at an assay dependent concentration.

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

    • Western blot - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
      Western blot - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
      Anti-alpha 1 Fetoprotein antibody [EPAFP61] (ab133617) at 1/10000 dilution (Purified) + HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 15 µg

      Secondary
      Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 68 kDa



      This data was developed using ab133617, the same antibody clone in a different buffer formulation.

    • Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
      Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
      This data was developed using ab133617, the same antibody clone in a different buffer formulation.
      Immunocytochemistry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling alpha 1 Fetoprotein with Purified ab133617 at 1/250 dilution (0.52 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
      This data was developed using ab133617, the same antibody clone in a different buffer formulation.
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver cancer tissue sections labeling alpha 1 Fetoprotein with Purified ab133617 at 1/100 dilution (1.3 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Flow Cytometry (Intracellular) - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
      Flow Cytometry (Intracellular) - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)

      This data was developed using ab133617, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of HepG2 Human hepatocellular carcinoma epithelial cell cells labeling alpha 1 Fetoprotein with purified ab133617 at 1/100 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue).

       

    • Immunoprecipitation - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
      Immunoprecipitation - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)

      This data was developed using ab133617, the same antibody clone in a different buffer formulation.
      Purified ab133617 at 1/20 dilution (0.5µg) immunoprecipitating alpha 1 Fetoprotein in HepG2 whole cell lysate.
      Lane 1 (input): HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab133617 + HepG2 whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133617 in HepG2 whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 70 kDa
      Lower bands could be caspase-dependent cleavage fragments. (PMID: 31811908)

    • Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)
      Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (ab215379)

    References (0)

    Publishing research using ab215379? Please let us know so that we can cite the reference in this datasheet.

    ab215379 has not yet been referenced specifically in any publications.

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