Human Pref-1/DLK1/FA1 Antibody

R&D Systems | Catalog # AF1144

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Mouse

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Pref-1/DLK1/FA1 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Pref-1 long isoform
Ala24-Pro297 with Arg248Pro and Lys295Ser substitutions
Accession # AAA75364

Specificity

Detects human Pref-1/DLK-1/FA1 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human Pref-1/DLK1/FA1 Antibody

Detection of Human Pref-1/DLK1/FA1 antibody by Western Blot.

Detection of Human Pref‑1/DLK1/FA1 by Western Blot.

Western blot shows lysates of human adrenal gland tissue and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Pref-1/DLK1/FA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1144) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Pref-1/DLK1/FA1 at approximately 45-60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Pref-1/DLK1/FA1 antibody in HepG2 Human Cell Line by Immunocytochemistry (ICC).

Pref‑1/DLK1/FA1 in HepG2 Human Cell Line.

Pref-1/DLK1/FA1 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Goat Anti-Human Pref-1/DLK1/FA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1144) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Mouse Pref-1/DLK1/FA1 by Western Blot

Detection of Mouse Pref-1/DLK1/FA1 by Western Blot

Dlk1 inhibits myoblast differentiation in vitro.A: C2C12 cells were constructed to constitutively express full-length Dlk1 under control by the CMV promotor, DLK1-C2C12. Cell morphology and myogenic differentiation potential was analyzed by immunocytochemistry. Forced expression of Dlk1 resulted in inhibition of myogenic differentiation observed as no formation of myotubes, lack of myostatin expression and reduced Desmin staining compared to C2C12 parental control cells. Addition of Dlk1 antibody to the culture supernatant during differentiation reverted the effect of forced Dlk1 expression observed as a regained ability to form myotubes and express myogenin compared to control cells. Scalebars: 20 µm in all images. B: DLK1-C2C12 cells (DLK1) and C2C12 parental control cells were cultured under proliferating conditions for 3 days followed by differentiation for another 5 days. Cells were harvested and analyzed at 1, 3, 5, and 8 days in culture. The differentiation process was analyzed by qPCR of Dlk1, the myogenic markers Mef2a, Myf5, Myod, and Myogenin in addition to Myostatin, Follistatin, TGF beta 1, Smad7 and Decorin. All qPCR analyses were run in triplicates on duplicate analyses containing triplicate samples (n = 2 for each time point) and presented as relative expression levels with normalization to.Actb, Gusb, Pgk1, Gapdh and Tfrc as described in materials and methods. Black bars: DLK1+ cells; white bars: control cells. C: Dlk1 (13 kDa band) and myostatin (a double band corresponding to 40–50 kDa) protein level was analyzed with western blotting during proliferation and differentiation of DLK1-C2C12 and control cells. Dlk1 protein was only expressed by DLK1-C2C12 cells and not by control cells. Myostatin protein was expressed in control cells during differentiation but not during proliferation of either of the cells lines or during differentiation of DLK1-C2C12 cells. However, addition of Dlk1 antibody resulted in expression of myostatin by the DLK1-C2C12 cells d
Detection of Mouse Pref-1/DLK1/FA1 by Immunocytochemistry/ Immunofluorescence

Detection of Mouse Pref-1/DLK1/FA1 by Immunocytochemistry/ Immunofluorescence

Dlk1 inhibits myoblast differentiation in vitro.A: C2C12 cells were constructed to constitutively express full-length Dlk1 under control by the CMV promotor, DLK1-C2C12. Cell morphology and myogenic differentiation potential was analyzed by immunocytochemistry. Forced expression of Dlk1 resulted in inhibition of myogenic differentiation observed as no formation of myotubes, lack of myostatin expression and reduced Desmin staining compared to C2C12 parental control cells. Addition of Dlk1 antibody to the culture supernatant during differentiation reverted the effect of forced Dlk1 expression observed as a regained ability to form myotubes and express myogenin compared to control cells. Scalebars: 20 µm in all images. B: DLK1-C2C12 cells (DLK1) and C2C12 parental control cells were cultured under proliferating conditions for 3 days followed by differentiation for another 5 days. Cells were harvested and analyzed at 1, 3, 5, and 8 days in culture. The differentiation process was analyzed by qPCR of Dlk1, the myogenic markers Mef2a, Myf5, Myod, and Myogenin in addition to Myostatin, Follistatin, TGF beta 1, Smad7 and Decorin. All qPCR analyses were run in triplicates on duplicate analyses containing triplicate samples (n = 2 for each time point) and presented as relative expression levels with normalization to.Actb, Gusb, Pgk1, Gapdh and Tfrc as described in materials and methods. Black bars: DLK1+ cells; white bars: control cells. C: Dlk1 (13 kDa band) and myostatin (a double band corresponding to 40–50 kDa) protein level was analyzed with western blotting during proliferation and differentiation of DLK1-C2C12 and control cells. Dlk1 protein was only expressed by DLK1-C2C12 cells and not by control cells. Myostatin protein was expressed in control cells during differentiation but not during proliferation of either of the cells lines or during differentiation of DLK1-C2C12 cells. However, addition of Dlk1 antibody resulted in expression of myostatin by the DLK1-C2C12 cells d
Detection of Mouse Pref-1/DLK1/FA1 by Immunocytochemistry/ Immunofluorescence

Detection of Mouse Pref-1/DLK1/FA1 by Immunocytochemistry/ Immunofluorescence

Dlk1 inhibits myoblast differentiation in vitro.A: C2C12 cells were constructed to constitutively express full-length Dlk1 under control by the CMV promotor, DLK1-C2C12. Cell morphology and myogenic differentiation potential was analyzed by immunocytochemistry. Forced expression of Dlk1 resulted in inhibition of myogenic differentiation observed as no formation of myotubes, lack of myostatin expression and reduced Desmin staining compared to C2C12 parental control cells. Addition of Dlk1 antibody to the culture supernatant during differentiation reverted the effect of forced Dlk1 expression observed as a regained ability to form myotubes and express myogenin compared to control cells. Scalebars: 20 µm in all images. B: DLK1-C2C12 cells (DLK1) and C2C12 parental control cells were cultured under proliferating conditions for 3 days followed by differentiation for another 5 days. Cells were harvested and analyzed at 1, 3, 5, and 8 days in culture. The differentiation process was analyzed by qPCR of Dlk1, the myogenic markers Mef2a, Myf5, Myod, and Myogenin in addition to Myostatin, Follistatin, TGF beta 1, Smad7 and Decorin. All qPCR analyses were run in triplicates on duplicate analyses containing triplicate samples (n = 2 for each time point) and presented as relative expression levels with normalization to.Actb, Gusb, Pgk1, Gapdh and Tfrc as described in materials and methods. Black bars: DLK1+ cells; white bars: control cells. C: Dlk1 (13 kDa band) and myostatin (a double band corresponding to 40–50 kDa) protein level was analyzed with western blotting during proliferation and differentiation of DLK1-C2C12 and control cells. Dlk1 protein was only expressed by DLK1-C2C12 cells and not by control cells. Myostatin protein was expressed in control cells during differentiation but not during proliferation of either of the cells lines or during differentiation of DLK1-C2C12 cells. However, addition of Dlk1 antibody resulted in expression of myostatin by the DLK1-C2C12 cells d

Applications for Human Pref-1/DLK1/FA1 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HepG2 human hepatocellular carcinoma cell line

Western Blot

1 µg/mL
Sample: Human adrenal gland tissue and human placenta tissue

Reviewed Applications

Read 1 review rated 5 using AF1144 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Pref-1/DLK1/FA1

Pref-1 (Preadipocyte factor 1; also DLK-1 and FA1) is a 58-65 kDa member of the Notch/Serrata/Delta family of proteins.  It is expressed in prechondrocytes and preadipocytes and appears to block progenitor cell differentiation into mature cell lineages.  Mature human Pref-1 is a 360 amino acid (aa) type I transmembrane N- and O-linked glycoprotein.  It contains a 280 aa extracellular region (aa 24-303), a 24 aa transmembrane segment (aa 304-327), and a 56 aa cytoplasmic domain (aa 328-383).  The extracellular region contains six EGF-like domains, and undergoes proteolytic cleavage to generate a bioactive 50 kDa fragment, plus three 25‑31 kDa fragments that show no activity.  There are multiple potential splice variants.  One shows a deletion of aa 229-301, a second possesses a six aa substitution for aa 1‑52, a third shows a deletion of aa 210-277, while a fourth contains a six aa substitution for aa # 207-383.  Over aa 24-297, human Pref-1 shares 82% aa identity with mouse Pref-1.            

Long Name

Preadipocyte Factor-1/Protein delta Homolog 1/Fetal Antigen 1

Alternate Names

DLK-1, DLK1, FA1, pG2, Pref1, ZOG

Entrez Gene IDs

8788 (Human); 13386 (Mouse)

Gene Symbol

DLK1

UniProt

AAA75364

Additional Pref-1/DLK1/FA1 Products

Product Documents for Human Pref-1/DLK1/FA1 Antibody

Certificate of Analysis

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Product Specific Notices for Human Pref-1/DLK1/FA1 Antibody

For research use only

Citations for Human Pref-1/DLK1/FA1 Antibody

Customer Reviews for Human Pref-1/DLK1/FA1 Antibody (1)

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  • Human Pref-1/DLK1/FA1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: NIH3T3 DLK1 overexpress whole cell lysate
    Species: Human
    Verified Customer | Posted 05/25/2018
    I tested several DLK1 antibodies produced mouse and rabbit, only this GOAT polyclonal antibody worked in western blot. There is almost no any background and nonspecific binding.
    Human Pref-1/DLK1/FA1 Antibody AF1144

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