rabbit anti cav3 1  (Alomone Labs)

Journal: International archives of allergy and immunology

Article Title: Expression of T-Type Voltage-Gated Calcium Channel in the Cilia of Human Nasal Epithelial Cells.

doi: 10.1159/000521765

Figure Lengend Snippet: Fig. 4. Western blot analysis. The collected turbinate mucosa was minced/ground and soaked in the RIPA lysis buffer with protease inhibitor and phosphatase inhibitor. The lysate was centrifuged, and the supernatant was mixed with EzApply and boiled. An ali- quot of the mixture equivalent to 10 μg protein was loaded onto the well of a 5–15% gradient polyacrylamide gel and electropho- resed at a constant current of 20 mA/gel for 80 min. The gel was then electrotransferred onto a nitrocellulose membrane at a con- stant current of 180 mA/gel for 60 min. The blotted membrane was incubated with rabbit anti-human Cav3.1 (CACNA1G) polyclonal antibody, rabbit anti-human Cav3.3 (CACNA1I) polyclonal anti- body, or mouse anti-human β-actin monoclonal antibody. The membrane was reacted with peroxidase-conjugated goat anti-rab- bit or anti-mouse IgG and soaked in Luminescence solutions for chemiluminescence detection. Images were captured by a Lu- miCube CMOS digital camera.

Article Snippet: The sections were hydrated in PBS with 0.3% Triton X-100 (PBST) for 20 min and treated with 1.5% normal goat serum in PBST for 1 h. They were then incubated with rabbit anti-human Cav3.1 (CACNA1G) polyclonal antibody (Cusabio, Houston, TX, USA), rabbit anti-human Cav3.2 (CACNA1H) polyclonal antibody (Novus Biologicals, Centennial, CO, USA), rabbit anti-human Cav3.3 (CACNA1I) D ow nloaded from http://karger.com /iaa/article-pdf/183/6/579/3762060/000521765.pdf by guest on 13 January 2025 T-Type VGCC in Nasal Mucosa 581Int Arch Allergy Immunol 2022;183:579–590 DOI: 10.1159/000521765 polyclonal antibody (Novus Biologicals), or rabbit anti-human Cav2.3 (CACNA1E) polyclonal antibody (Lifespan BioSciences, Seattle, WA, USA) at 4°C overnight.

Techniques: Western Blot, Lysis, Protease Inhibitor, Membrane, Incubation

Journal: International archives of allergy and immunology

Article Title: Expression of T-Type Voltage-Gated Calcium Channel in the Cilia of Human Nasal Epithelial Cells.

doi: 10.1159/000521765

Figure Lengend Snippet: Fig. 5. Expressions of CACNA1G mRNA and CACNA1I mRNA. The collected turbinate mucosa was minced and soaked in TRIzol reagent. Total RNA was extracted by the acid guanidiniumthiocy- anate-phenol-chloroform method, cleaned up with a BioRobot EZ1 system, and reverse-transcribed to cDNA using a High-Ca- pacity RNA-to-cDNA Kit. The real-time reverse transcription- polymerase chain reaction analysis was performed with an Applied Biosystems StepOnePlus real-time PCR system using TaqMan Fast Universal PCR Master Mix with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a housekeeping gene. The thermal cy- cler conditions were as follows: holding at 95°C for 2 min, followed

Article Snippet: The sections were hydrated in PBS with 0.3% Triton X-100 (PBST) for 20 min and treated with 1.5% normal goat serum in PBST for 1 h. They were then incubated with rabbit anti-human Cav3.1 (CACNA1G) polyclonal antibody (Cusabio, Houston, TX, USA), rabbit anti-human Cav3.2 (CACNA1H) polyclonal antibody (Novus Biologicals, Centennial, CO, USA), rabbit anti-human Cav3.3 (CACNA1I) D ow nloaded from http://karger.com /iaa/article-pdf/183/6/579/3762060/000521765.pdf by guest on 13 January 2025 T-Type VGCC in Nasal Mucosa 581Int Arch Allergy Immunol 2022;183:579–590 DOI: 10.1159/000521765 polyclonal antibody (Novus Biologicals), or rabbit anti-human Cav2.3 (CACNA1E) polyclonal antibody (Lifespan BioSciences, Seattle, WA, USA) at 4°C overnight.

Techniques: Reverse Transcription, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Journal: International archives of allergy and immunology

Article Title: Expression of T-Type Voltage-Gated Calcium Channel in the Cilia of Human Nasal Epithelial Cells.

doi: 10.1159/000521765

Figure Lengend Snippet: Fig. 4. Western blot analysis. The collected turbinate mucosa was minced/ground and soaked in the RIPA lysis buffer with protease inhibitor and phosphatase inhibitor. The lysate was centrifuged, and the supernatant was mixed with EzApply and boiled. An ali- quot of the mixture equivalent to 10 μg protein was loaded onto the well of a 5–15% gradient polyacrylamide gel and electropho- resed at a constant current of 20 mA/gel for 80 min. The gel was then electrotransferred onto a nitrocellulose membrane at a con- stant current of 180 mA/gel for 60 min. The blotted membrane was incubated with rabbit anti-human Cav3.1 (CACNA1G) polyclonal antibody, rabbit anti-human Cav3.3 (CACNA1I) polyclonal anti- body, or mouse anti-human β-actin monoclonal antibody. The membrane was reacted with peroxidase-conjugated goat anti-rab- bit or anti-mouse IgG and soaked in Luminescence solutions for chemiluminescence detection. Images were captured by a Lu- miCube CMOS digital camera.

Article Snippet: The sections were then treated with 1% normal BSA in PBST for 1 h and incubated with rabbit antihuman Cav3.1 (CACNA1G) polyclonal antibody (Cusabio) or rabbit anti-human Cav3.3 (CACNA1I) polyclonal antibody (Novus Biologicals) at 4°C overnight.

Techniques: Western Blot, Lysis, Protease Inhibitor, Membrane, Incubation

Journal: International archives of allergy and immunology

Article Title: Expression of T-Type Voltage-Gated Calcium Channel in the Cilia of Human Nasal Epithelial Cells.

doi: 10.1159/000521765

Figure Lengend Snippet: Fig. 5. Expressions of CACNA1G mRNA and CACNA1I mRNA. The collected turbinate mucosa was minced and soaked in TRIzol reagent. Total RNA was extracted by the acid guanidiniumthiocy- anate-phenol-chloroform method, cleaned up with a BioRobot EZ1 system, and reverse-transcribed to cDNA using a High-Ca- pacity RNA-to-cDNA Kit. The real-time reverse transcription- polymerase chain reaction analysis was performed with an Applied Biosystems StepOnePlus real-time PCR system using TaqMan Fast Universal PCR Master Mix with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a housekeeping gene. The thermal cy- cler conditions were as follows: holding at 95°C for 2 min, followed

Article Snippet: The sections were then treated with 1% normal BSA in PBST for 1 h and incubated with rabbit antihuman Cav3.1 (CACNA1G) polyclonal antibody (Cusabio) or rabbit anti-human Cav3.3 (CACNA1I) polyclonal antibody (Novus Biologicals) at 4°C overnight.

Techniques: Reverse Transcription, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction