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  1. Link

    fgf21-antibody-epr83142-ab171941.pdf

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Cardiovascular Angiogenesis Growth Factors FGF FGF
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • SDS
Reviews (1) Submit a question References (55)

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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

You may also be interested in

Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Primary
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Anti-PGC1 alpha + beta antibody [EPR18289] - BSA and Azide free (ab250943)
Protein
Recombinant human FGF21 protein (Active) (ab54141)

View more associated products

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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Allow to warm to room temp and agitate gently before aliquotting. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR8314(2)
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Growth Factors
    • FGF
    • FGF
    • Signal Transduction
    • Growth Factors/Hormones
    • FGF
    • Cancer
    • Growth factors
    • FGF

Associated products

  • Alternative Versions

    • Anti-FGF21 antibody [EPR8314(2)] - Low endotoxin, Azide free (ab219368)
    • Anti-FGF21 antibody [EPR8314(2)] - BSA and Azide free (ab271925)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human FGF21 knockout HeLa cell line (ab265974)
  • KO cell lysates

    • Human FGF21 knockout HeLa cell lysate (ab256915)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
  • Recombinant Protein

    • Recombinant human FGF21 protein (Active) (ab54141)

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

  • Function

    Stimulates glucose uptake in differentiated adipocytes via the induction of glucose transporter SLC2A1/GLUT1 expression (but not SLC2A4/GLUT4 expression). Activity requires the presence of KLB.
  • Sequence similarities

    Belongs to the heparin-binding growth factors family.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession Q9NSA1 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 26291 Human
    • Entrez Gene: 56636 Mouse
    • Entrez Gene: 170580 Rat
    • Omim: 609436 Human
    • SwissProt: Q9NSA1 Human
    • SwissProt: Q9JJN1 Mouse
    • Unigene: 283015 Human
    • Unigene: 143736 Mouse
    • Alternative names

      • FGF 21 antibody
      • FGF-21 antibody
      • Fgf21 antibody
      • FGF21_HUMAN antibody
      • FGFL antibody
      • Fibroblast growth factor 21 antibody
      • PRO10196 antibody
      • UNQ3115 antibody
      see all

    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)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    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

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

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

    Average
    Abreviews
    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
    Read More

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    Verified customer

    Submitted Jun 25 2020

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