32-6000

antibody from Invitrogen Antibodies

Targeting: DSG1 CDHF4, DSG

Provider product page for 32-6000

Western blot

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [19]
Comments [0]
Validations
Immunohistochemistry [2]
Other assay [5]

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Product number: 32-6000 - Provider product page
Provider: Invitrogen Antibodies
Product name: Desmoglein 1 Monoclonal Antibody (27B2)
Antibody type: Monoclonal
Antigen: Other
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 27B2
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: -20°C

DSG1 protein structure - 32-6000 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Desmoglein-1 showing staining in the cytoplasm and membrane of paraffin-embedded human skin tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Desmoglein-1 monoclonal antibody (Product # 32-6000) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Desmoglein-1 showing staining in the cytoplasm and membrane of paraffin-embedded human skin tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Desmoglein-1 monoclonal antibody (Product # 32-6000) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3. Ligand targeting of EphA2 triggers keratinocyte colony compaction, stratification and differentiation. (A) Phalloidin (F-actin; in green) and DAPI (nuclei; in blue) staining of sparse keratinocytes that were treated with 1.0 mug/ml Fc or ephrin-A1-Fc for 7 d in 1.2 mM calcium (scale bar, 50 mum). (B) Cell compaction was analyzed by using ImageJ software to measure the surface area of individual cells and presented in a box-and-whisker plot. (C) Maximal image projections (top panels) from peptide-treated colonies that were stained with phalloidin to image F-actin (in green), an antibody against keratin 10 (in red) or DAPI to visualize nuclei (in blue). Apotome-processed images were acquired from stratified keratinocyte colonies at 0.25 mum increments (scale bar, 50 mum). Images below depict cross-sections through the z-x axis that were magnified in the z plane to better resolve keratinocyte piles (scale bar, 10 mum). Colonies that had formed in the presence of Fc or ephrin-A1-Fc peptide in 1.2 mM calcium were also fixed and immunostained for desmocollin 1 a/b (in green) or keratin 10 (in red) to detect stratified keratinocytes (scale bar, 100 mum). (D) siCtrl or siEphA2 keratinocytes were seeded under sparse conditions and treated with 0.01, 0.1, or 1.0 mug/ml Fc or ephrin-A1-Fc peptide for 7 days in 1.2 mM calcium. Western blot analysis of lysates prepared from these cultures were probed for EphA2, desmoglein 1, desmocollin 1 a/b, desmoglein 3, E-cadherin, keratin 1

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5. Ligand targeting of EphA2 enhances keratinocyte adhesion and differentiation in a desmoglein-1 dependent manner. (A) Confluent keratinocyte sheets were treated with 1.0 mug/ml Fc or ephrin-A1-Fc peptide for 48 h in 0.4 mM calcium and incubated in the presence of 2.0 mug/ml recombinant Staphylococcus aureus exfoliative toxin A (ETA) for an additional 2 h. Top panels show Western blot analysis of EphA2, full length (FL) and cleaved (CL) desmoglein 1, desmocollin 1 a/b, desmoglein 3, E-cadherin, or GAPDH. A mechanical dissociation assay was performed, and the bottom panel contains a bar graph depicting the average number of fragments. (B) Keratinocytes transduced with retroviruses harboring microRNA mimetic sequences targeting lamin A/C (mirLmn) as a control or desmoglein 1 (mirDsg1) were seeded to confluency and treated with 1.0 mug/ml ephrin-A1-Fc peptide for 48 h in 0.4 mM calcium. Top panels show Western blot analysis of EphA2, desmoglein 1, desmocollin 1 a/b, desmoglein 3, E-cadherin, or GAPDH. In parallel, a mechanical dissociation assay was performed (bottom panel). (C) Western blot analysis for EphA2, desmoglein 1, desmocollin 1 a/b, desmoglein 3, E-cadherin, keratin 10, or GAPDH from confluent keratinocytes transduced with mirLmn and mirDsg1 that were treated with 1.0 mug/ml Fc or ephrin-A1-Fc peptide for 48 h in 1.2 mM calcium. (D) Keratinocytes transduced with mirLmn and mirDsg1 were seeded under sparse conditions and treated with 0.01, 0.1, or 1.0 mug/ml Fc

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 8 Fluorescent photomicrographs of cultured progenitor cells (P3) from unaffected (A,B) and laminitic (C,D) hooves labeled (red) with anti- DSG1 (A,C) , DSG3 (B,D) antibodies or no antibodies (E) . DAPI nuclear stain (blue); Magnification = 40X (A-D) ; 20X (E) .

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 9 Representative immunofluorescent photomicrographs showing the protein expression levels of differentiation (filaggrin) and desmosomal (desmoglein-1) protein markers in control RHSE and FTRHSP under different treatment conditions versus fisetin or Vit-D 3 -treated FTRHSP tissues. Results are representative of three independent experiments each performed in quadruplicate and comparing control RHSE vs. FTRHSP treated groups. Data green (Dsgl-1); red (filaggrin), blue (DAPI) and mixed is merged representation. Dsgl-1 = desmoglein-1. Scale bar = 20 mum.