Revision 1
Cell Signaling Technology

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Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
AFP (D12C1) Rabbit mAb 4448 40 µl 65 kDa Rabbit IgG
N-Cadherin Antibody 4061 40 µl 140 kDa Rabbit 
EOMES Antibody 4540 40 µl 70 kDa Rabbit 
FoxA2/HNF3β (D56D6) XP® Rabbit mAb 8186 40 µl 50 kDa Rabbit IgG
GATA-6 (D61E4) XP® Rabbit mAb 5851 40 µl 55 kDa Rabbit IgG
HNF4α (C11F12) Rabbit mAb 3113 40 µl 52 kDa Rabbit IgG
PDGF Receptor α (D13C6) XP® Rabbit mAb 5241 40 µl 190 kDa Rabbit IgG
Sall4 (D16H12) Rabbit mAb 8459 40 µl 80, 142 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Endodermal Lineage Marker Antibody Sampler Kit provides an economical means of evaluating proteins expressed during endoderm development. This kit contains enough antibody to perform four western blot experiments per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Two endodermal lineages develop during mammalian embryogenesis, the primitive endoderm of the blastocyst stage embryo and the definitive endoderm at gastrulation. The primitive endoderm gives rise to extra-embryonic lineages encompassing the visceral and the parietal endoderm. The definitive endoderm contributes to the respiratory and gastrointestinal tracts by forming the epithelial lining of the trachea, esophagus, lungs, stomach and intestines, and is a major component of many glands, including thyroid, thymus, pancreas and liver (1). Understanding molecular mechanisms that regulate early endodermal fates is seminal for the advance of stem cell research as they connect the transition from pluripotency to endoderm specification during mammalian development and contribute to the generation of clinically relevant cell types. FoxA2/HNF3β is a transcription factor essential for development of the endoderm and midline structures in mouse embryos (2,3). EOMES acts during gastrulation to promote the specification of the definitive endoderm (4). Markers of hepatic differentiation in the endoderm include expression of α-fetoprotein (AFP) and N-cadherin (5,6). HNF4α is involved in the differentiation of the visceral endoderm. GATA-6 lies upstream of HNF4 in a transcriptional cascade that regulates differentiation of the visceral endoderm and is also required for the establishment of the endodermally derived bronchial epithelium (7). Sall4 is required for the formation of the primitive endoderm from inner cell mass. It has been reported that extra-embryonic stem cell lines cannot be formed in Sall4-deficient blastocysts (8). PDGF receptor α is expressed in primitive endoderm derivatives throughout embryogenesis (9).

  1. Wells, J.M. and Melton, D.A. (1999) Annu Rev Cell Dev Biol 15, 393-410.
  2. Weinstein, D.C. et al. (1994) Cell 78, 575-88.
  3. Ang, S.L. and Rossant, J. (1994) Cell 78, 561-74.
  4. Costello, I. et al. (2011) Nat Cell Biol 13, 1084-91.
  5. Zhao, D. et al. (2009) PLoS One 4, e6468.
  6. Meier, V. et al. (2006) Comp Hepatol 5, 2.
  7. Morrisey, E.E. et al. (1998) Genes Dev 12, 3579-90.
  8. Elling, U. et al. (2006) Proc Natl Acad Sci U S A 103, 16319-24.
  9. Orr-Urtreger, A. et al. (1992) Development 115, 289-303.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #12693

    Endodermal Lineage Marker Antibody Sampler Kit

    Endodermal Lineage Marker Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extract from HepG2 cells using HNF4α (C11F12) Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from A431, MCF-7, C2C12, HUVEC, BAEC, C6 and H-4-II-E cells, using N-cadherin Antibody.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from Hep G2 cells using AFP (D12C1) Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from E6.5 and E10.5 stage mouse embryos using EOMES Antibody.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from various cell lines using PDGF Receptor α (D13C6) XP® Rabbit mAb (upper), PDGF Receptor β (28E1) Rabbit mAb #3169 (middle), and β-Actin Antibody #4967 (lower).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from Huh7 and 293 cells using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 7 Expand Image
    CUT&Tag was performed with Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across HNMT, a known target gene of GATA-6 (see our CUT&RUN-qPCR figure).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 9 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from Hep G2 cells using FoxA2/HNF3β (D56D6) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from NCCIT cells using Sall4 (D16H12) Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin embedded human hepatocellular carcinoma using HNF4α (C11F12) Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 13 Expand Image
    Immunohistochemical analysis of paraffin-embedded human astrocytoma using PDGF Receptor α (D13C6) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of KM12 (left) and SK-OV-3 cells (right) using GATA-6 (D61E4) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 15 Expand Image
    CUT&Tag was performed with Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 2 (upper), including HNMT (lower), a known target gene of GATA-6 (see our CUT&RUN-qPCR figure).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 16 Expand Image
    Immunohistochemical analysis of paraffin-embedded human granulosa cell tumor of the ovary using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 17 Expand Image
    Confocal immunofluorescent analysis of Hep G2 (left) and HeLa (right) cells using FoxA2/HNF3β (D56D6) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 18 Expand Image
    Confocal immunofluorescent analysis of NTERA-2 (left) and HeLa cells (right) using Sall4 (D16H12) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded HepG2 cells in the presence of control peptide (left) or antigen specific peptide (right) using HNF4α (C11F12E8) Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded NCI-H1703 (PDGFRα+, left) and HCC827 (PDGFRα-, right) cell pellets using PDGF Receptor α (D13C6) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 21 Expand Image
    Flow cytometric analysis of SKOV3 cells (blue) and HUH-7 cells (green) using GATA-6 (D61E4) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded human gastric carcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 23 Expand Image
    Confocal immunofluorescent analysis of HepG2 (left) and HeLa cells (right) using HNF4α (C11F12) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded human leiomyosarcoma using PDGF Receptor α (D13C6) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 25 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across FGFR2, a known target gene of GATA-6 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 27 Expand Image
    Confocal immunofluorescent analysis of NCI-H1703 (left), A172 (center) and HCC827 cells (right) using PDGF Receptor α (D13C6) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 28 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 10 (upper), including FGFR2 (lower), a known target gene of GATA-6 (see additional figure containing ChIP-qPCR data).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 30 Expand Image
    Flow cytometric analysis of A-204 cells using PDGF Receptor α (D13C6) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 31 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from Caco-2 cells and either GATA-6 (D61E4) XP® Rabbit mAb, or Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human FDPS Promoter Primers #13840, human FGFR2 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 33 Expand Image
    CUT&RUN was performed with Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across FGFR2 gene.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 35 Expand Image
    CUT&RUN was performed with Caco-2 cells and GATA-6 (D61E4) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 10 (upper), including FGFR2 gene (lower).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human stomach using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 37 Expand Image
    CUT&RUN was performed with Caco-2 cells and either GATA-6 (D61E4) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human HNMT upstream primer, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human small intestine using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 39 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human pancreas using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 40 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human ovary using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 41 Expand Image
    >Immunohistochemical analysis of paraffin-embedded normal human lung using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 42 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human heart using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 43 Expand Image
    Immunohistochemical analysis of paraffin-embedded human appendix using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 44 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human adrenal gland using GATA-6 (D61E4) XP ® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 45 Expand Image
    Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 46 Expand Image
    Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 47 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse testis using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 48 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse ovary using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 49 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse colon using GATA-6 (D61E4) XP® Rabbit mAb
    Endodermal Lineage Marker Antibody Sampler Kit: Image 50 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse small intestine using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 51 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using GATA-6 (D61E4) XP® Rabbit mAb.
    Endodermal Lineage Marker Antibody Sampler Kit: Image 52 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human colon using GATA-6 (D61E4) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    Endodermal Lineage Marker Antibody Sampler Kit: Image 53 Expand Image

    Immunohistochemical analysis of paraffin-embedded HuH-7 cell pellet (left, positive) and SUP-B15 cell pellet (right, negative) using GATA-6 (D61E4) XP® Rabbit mAb.