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13236
Adherens Junction Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Adherens Junction Antibody Sampler Kit #13236

Citations (0)
Adherens Junction Antibody Sampler Kit: Image 1
Western blot analysis of extracts from various cell lines using Afadin (D1Y3Z) Rabbit mAb.
Adherens Junction Antibody Sampler Kit: Image 2
Immunoprecipitation of afadin from A-431 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Afadin (D1Y3Z) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Afadin (D1Y3Z) Rabbit mAb.
Adherens Junction Antibody Sampler Kit: Image 3
Western blot analysis of extracts from 293, MCF-7 and C2C12 cells using γ-Catenin Antibody.
Adherens Junction Antibody Sampler Kit: Image 4
Immunohistochemical analysis of paraffin-embedded human breast carcioma using gamma-Catenin Antibody.
Adherens Junction Antibody Sampler Kit: Image 5
Immunohistochemical analysis of paraffin-embedded human colon carcioma using gamma-Catenin Antibody.
Adherens Junction Antibody Sampler Kit: Image 6
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using gamma-Catenin Antibody.
Adherens Junction Antibody Sampler Kit: Image 7
Confocal immunofluorescent analysis of HT-29 cells labeled with gamma-Catenin Antibody (green). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
Adherens Junction Antibody Sampler Kit: Image 8
Western blot analysis of extracts from HUVEC, NIH/3T3, MCF-7, ZR-75 and A431 cells using α-E-Catenin (23B2) Rabbit mAb.
Adherens Junction Antibody Sampler Kit: Image 9
Western blot analysis of extracts from MDA-MB-231 and NIH/3T3 cells using Catenin δ-1 Antibody.
Adherens Junction Antibody Sampler Kit: Image 10
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.
Adherens Junction Antibody Sampler Kit: Image 11
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and either β-Catenin (D10A8) XP® Rabbit mAb or Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across AXIN2, a known target gene of β-Catenin (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Adherens Junction Antibody Sampler Kit: Image 12
Confocal immunofluorescent analysis of HeLa (left) and NCI-H28 (right) cells using β-Catenin (D10A8) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Adherens Junction Antibody Sampler Kit: Image 13
Western blot analysis of extracts from control HeLa cells (lane 1) or HeLa cells with an apparent in-frame truncation mutation in the gene encoding β-Catenin (lane 2) using β-Catenin (D10A8) XP® Rabbit mAb, #8480 (upper) or β-actin (D6A8) Rabbit mAb #8457 (lower). The change in β-Catenin molecular weight in the mutated HeLa cells is consistent with an in-frame deletion.
Adherens Junction Antibody Sampler Kit: Image 14
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and either β-Catenin (D10A8) XP® Rabbit mAb or Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 17 (upper), including AXIN2 (lower), a known target gene of β-Catenin (see additional figure containing ChIP-qPCR data).
Adherens Junction Antibody Sampler Kit: Image 15
Western blot analysis of extracts from various cell lines using β-Catenin (D10A8) XP® Rabbit mAb.
Adherens Junction Antibody Sampler Kit: Image 16
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and either β-Catenin (D10A8) 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 Axin2 Intron 1 Primers #8973, SimpleChIP® Human CaMK2D Intron 3 Primers #5111, human c-Myc 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.
Adherens Junction Antibody Sampler Kit: Image 17
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.
Adherens Junction Antibody Sampler Kit: Image 18
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.
Adherens Junction Antibody Sampler Kit: Image 19
Immunohistochemical analysis of paraffin-embedded cell pellets, HeLa (left) or NCI-H28 (right), using β-Catenin (D10A8) XP® Rabbit mAb.
Adherens Junction Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded mouse colon using β-Catenin (D10A8) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Adherens Junction Antibody Sampler Kit: Image 21
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using ß-Catenin (D10A8) XP® Rabbit mAb.

Adherens Junction Antibody Sampler Kit: Image 22
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using ß-Catenin (D10A8) XP® Rabbit mAb.
Adherens Junction Antibody Sampler Kit: Image 23
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.
Adherens Junction Antibody Sampler Kit: Image 24
Confocal immunofluorescent analysis of mouse colon using β-Catenin (D10A8) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Adherens Junction Antibody Sampler Kit: Image 25
Flow cytometric analysis of NCI-H28 cells (blue) and HeLa cells (green) using β-Catenin (D10A8) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as secondary antibody.
Inquiry Info.# 13236

Product Description

The Adherens Junction Antibody Sampler Kit provides an economical means of detecting the protein components of adherens junctions. The kit includes enough antibody to perform four western blot experiments per primary antibody.

Specificity / Sensitivity

Each antibody in this kit detects endogenous levels of total protein for the specified target and does not cross-react with other family members unless indicated. The α-E-Catenin (23B2) Rabbit mAb may cross-react with neuronal α-N-catenin. The γ-Catenin Antibody does not cross-react with β-catenin. Based on sequence homology, the Afadin (D1Y3Z) Rabbit mAb is expected to recognize all isoforms of afadin.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of human γ-catenin protein, or the sequence of human catenin δ-1 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino-terminal sequence of human α-E-catenin, Arg1117 of human afadin protein, or to residues surrounding Pro714 of human β-catenin protein.

Background

Adherens junctions are dynamic structures that form cell-cell contacts and are important in development, differentiation, tissue integrity, morphology and cell polarity. They are composed of the transmembrane proteins, cadherins, which bind cadherins on adjacent cells in a calcium-dependent manner. On the cytoplasmic side of adherens junctions, the classic model states that cadherins are linked to the cytoskeleton through β- and α-catenin. α-E-catenin is ubiquitously expressed, α-N-catenin is expressed in neuronal tissue, and α-T-catenin is primarily expressed in heart tissue. Research studies have demonstrated that loss of E-cadherin and α-E-catenin occurs during the progression of several human cancers, indicating that the breakdown of adherens junctions is important in cancer progression (reviewed in 1). Research studies also suggest that, rather than acting as a static link between cadherins and actin, α-catenin regulates actin dynamics directly, possibly by competing with the actin nucleating arp2/3 complex (2,3). α-catenin also plays a role in regulating β-catenin-dependent transcriptional activity, affecting differentiation and response to Wnt signaling. α-catenin binds to β-catenin in the nucleus, preventing it from regulating transcription, and levels of both proteins appear to be regulated via proteasome-dependent degradation (4). Afadin has two splice variants: l-afadin, which is ubiquitously expressed, and s-afadin, which is expressed predominantly in neural tissue. s-afadin is a shorter form lacking one of the three proline-rich regions found in l-afadin, as well as the carboxyl-terminal F-actin binding region (5). Human s-afadin is identical to AF-6, the ALL-1 fusion partner involved in acute myeloid leukemias (6). Recent research has also shown that afadin is involved in controlling the directionality of cell movement when it is localized at the leading edge of moving cells (7,8).
  1. Kobielak, A. and Fuchs, E. (2004) Nat Rev Mol Cell Biol 5, 614-25.
  2. Yamada, S. et al. (2005) Cell 123, 889-901.
  3. Drees, F. et al. (2005) Cell 123, 903-15.
  4. Hwang, S.G. et al. (2005) J Biol Chem 280, 12758-65.
  5. Mandai, K. et al. (1997) J Cell Biol 139, 517-28.
  6. Prasad, R. et al. (1993) Cancer Res 53, 5624-8.
  7. Miyata, M. et al. (2009) J Cell Sci 122, 4319-29.
  8. Miyata, M. et al. (2009) J Biol Chem 284, 24595-609.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.
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