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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-FGF21 antibody [EPR8314(2)] (ab171941)

Reviews (1) Submit a question References (55)
Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
  • Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
  • Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
  • Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
  • Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
  • Anti-FGF21 antibody [EPR8314(2)] (ab171941)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR8314(2)] to FGF21
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-FGF21 antibody [EPR8314(2)]
    See all FGF21 primary antibodies

  • Description

    Rabbit monoclonal [EPR8314(2)] to FGF21

  • Host species

    Rabbit

  • Specificity

    The immunogen used for this product shares 6 continuous identical amino acids with SIKE1. Cross-reactivity with this protein has not been confirmed experimentally.

    Expression levels of the target protein vary with sample type and some optimisation may be required (PMID: 27285327).  For western blot using cell lines, it may be necessary to collect cell culture supernatant for endogenous FGF21 detection as the target protein is readily secreted (PMID: 24041694; PMID: 26691139).

  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human FGF21 aa 1-100. The exact sequence is proprietary.
    Database link: Q9NSA1

  • Positive control

    • Recombinant human FGF21 + IgG1 fusion protein (Fc Chimera Active) (ab108556) can be used as a positive control in WB. WB: Human fetal liver lysate, mouse spleen, rat spleen. IHC-P: Human stomach

  • General notes

    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.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab171941 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
WB (1)
1/1000. Predicted molecular weight: 22 kDa.
IHC-P
1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
Notes
WB
1/1000. Predicted molecular weight: 22 kDa.
IHC-P
1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
Application notes
Is unsuitable for ICC/IF.

Target

Images

  • Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    All lanes : Anti-FGF21 antibody [EPR8314(2)] (ab171941) at 1/1000 dilution

    Lane 1 : Human Liver cell lysate
    Lane 2 : Mouse Spleen cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 22 kDa
    Observed band size: 21 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-FGF21 antibody [EPR8314(2)] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab171941 was shown to bind specifically to FGF21. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    All lanes : Anti-FGF21 antibody [EPR8314(2)] (ab171941) at 1/1000 dilution

    Lane 1 : Wild-type HeLa Treated BFA (5 ug/mL, 6 h) cell lysate at 20 µg
    Lane 2 : FGF21 knockout HeLa Treated BFA (5 ug/mL, 6 h) cell lysate at 20 µg
    Lane 3 : Wild-type HeLa Vehicle Control BFA (0 ug/mL, 6 h) cell lysate at 20 µg
    Lane 4 : FGF21 knockout HeLa Vehicle Control BFA (0 ug/mL, 6 h) cell lysate at 20 µg
    Lane 5 : HepG2 Treated BFA (5 ug/mL, 6 h) cell lysate at 20 µg
    Lane 6 : HepG2 cell lysate at 20 µg
    Lane 7 : Empty
    Lane 8 : Human Liver cell lysate at 10 µg
    Lane 9 : Mouse Spleen cell lysate at 10 µg

    Performed under reducing conditions.

    Predicted band size: 22 kDa
    Observed band size: 21 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-FGF21 antibody [EPR8314(2)] staining at 1/1000 dilution, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab171941 was shown to bind specifically to FGF21. A band was observed at 21 kDa in treated wild-type HeLa cell lysates with no signal observed at this size in FGF21 knockout cell line ab265974 (knockout cell lysate ab256915). To generate this image, wild-type and FGF21 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development and with high-sensitivity chemiluminescence substrate and imaged with 16 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Anti-FGF21 antibody [EPR8314(2)] (ab171941) at 1/1000 dilution (purified) + Mouse spleen tissue lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 22 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Anti-FGF21 antibody [EPR8314(2)] (ab171941) at 1/5000 dilution (purified) + Rat spleen tissue lysate at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 22 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Anti-FGF21 antibody [EPR8314(2)] (ab171941) at 1/5000 dilution (purified) + Human fetal liver tissue lysate at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 22 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Immunohistochemical staining of paraffin embedded human stomach with purified ab171941 at a working dilution of 1/250. The secondary antibody used is ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Anti-FGF21 antibody [EPR8314(2)] (ab171941)
    Anti-FGF21 antibody [EPR8314(2)] (ab171941)

References (55)

Publishing research using ab171941? Please let us know so that we can cite the reference in this datasheet.

ab171941 has been referenced in 55 publications.

  • Chen X  et al. PPARa agonist WY-14,643 enhances ethanol metabolism in mice: Role of catalase. Free Radic Biol Med 169:283-293 (2021). PubMed: 33892114
  • Zhang X  et al. MiniCAFE, a CRISPR/Cas9-based compact and potent transcriptional activator, elicits gene expression in vivo. Nucleic Acids Res 49:4171-4185 (2021). PubMed: 33751124
  • Dai H  et al. FGF21 facilitates autophagy in prostate cancer cells by inhibiting the PI3K-Akt-mTOR signaling pathway. Cell Death Dis 12:303 (2021). PubMed: 33753729
  • Xu H  et al. Nuciferine Inhibited the Differentiation and Lipid Accumulation of 3T3-L1 Preadipocytes by Regulating the Expression of Lipogenic Genes and Adipokines. Front Pharmacol 12:632236 (2021). PubMed: 33841152
  • Athinarayanan S  et al. Fatty Acid Desaturase 1 Influences Hepatic Lipid Homeostasis by Modulating the PPARa-FGF21 Axis. Hepatol Commun 5:461-477 (2021). PubMed: 33681679
View all Publications for this product

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Western blot abreview for Anti-FGF21 antibody [EPR8314(2)]

 Average
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (heart)
Gel Running Conditions
Non-reduced Denaturing (15%)
Loading amount
30 µg
Specification
heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Verified customer

Submitted Jun 25 2020

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