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Western blot analysis of Desmoglein 3 on different lysates with Rabbit anti-Desmoglein 3 antibody (ER1901-81) at 1/40,000 dilution.
Lane 1: HaCaT (Human immortalized keratinocyte) cell lysate
Lane 2: A431 (Human epidermoid carcinoma skin squamous cell) cell lysate
Lysates/proteins at 15 µg/Lane.
Exposure time: 25 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: ER1901-81, 1/40,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 108 kDa
Observed band size: 130 kDa
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Application: Immunocytochemistry (IF-cell)
Species: Human
Sample: A431 (Human epidermoid carcinoma skin squamous cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: ER1901-81, 1/200, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
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Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Esophagus
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ER1901-81, 1/800, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Esophagus
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ER1901-81, 1/800, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Flow Cytometry (Intra)
Species: Human
Sample: A431 (Human epidermoid carcinoma skin squamous cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: ER1901-81 (1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"