- MA1-19732 - Invitrogen Antibodies PRKACA antibody

No submitted references: Submit reference

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of human protein kinase A catalytic (PKAc) alpha subunit was performed by loading 40 µg of Jlat-full whole cell lysate in reducing sample buffer onto a 4-15% tris-glycine gel. Proteins were transferred to PVDF and then blocked in blocking buffer (TBST+5% non-fat milk) for one hour at room temperature. PKC alpha was detected at approximately 42 kDa using a PKA alpha Monoclonal Antibody (6D2.1) (Product # MA1-19732) at a dilution of 1:500 overnight at 4°C on a rocking platform, followed by a goat anti-mouse IgG Horseradish Peroxidase-conjugated Antibody at a dilution of 1:2, 500 for one hour at room temperature. Chemiluminescent detection was performed.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: One million bone marrow-derived macrophages were dispensed into wells of a 48-well tissue culture plate and incubated with medium alone or with 5 x106 Histoplasma capsulatum yeast cells or with 300 mg/mL of cobalt chloride (CoCl2) for 24 h at 37°C in 5% CO2. Western blot analysis of HIF-2 alpha was performed by loading 150 µg whole cell lysate in reducing sample buffer onto a 4-20% gradient acrylamide gel. Proteins were transferred to nitrocellulose.HIF-2 alpha was detected at approximately 97 kDa using a HRP conjugated HIF-2 alpha Monoclonal Antibody (ep190b) (Product # MA1-19732) at a dilution of 1:500 overnight at 4°C. Chemiluminescent detection was performed.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using anti-PKA alpha Monoclonal Antibody (6D2.1) (Product # MA1-19732) and a 40 kDa band corresponding to PRKACA was observed across cell lines tested along with an uncharacterized band at ~60kDa. Whole cell extracts (30 µg lysate) of NIH: OVCAR-3 (Lane 1), A549 (Lane 2) Jurkat (Lane 3) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (MA1-19732) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Dot Blot analysis of GST and GST-fusion proteins using a nitrocellulose membrane (5 ng/spot); Lane 1: GST-Akt1, Lane 2: GST-Akt2, Lane 3: GST-Akt3, Lane 4: GST-PKAc alpha, Lane 5: GST-PKAc beta, Lane 6: GST-PKAc gamma, Lane 7: GST-MEK 1, Lane 8: GST.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Dot blot analysis of GST and GST-fusion proteins using anti-PKAc (6D2.1) and anti-GST (S-tag-05). The total amount of material spotted on the nitrocellulose membrane is 5 ng/spot. Lane 1: GST-Akt1; Lane 2: GST-Akt2; Lane 3: GST-Akt3; Lane 4: GST-PKAc alpha Monoclonal antibody (Product # MA1-19732); Lane 5: GST-PKAc beta; Lane 6: GST-PKAc gamma; Lane 7: GST-MEK 1; Lane 8: GST.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Dot blot analysis of GST and GST-fusion proteins using anti-PKAc (6D2.1) and anti-GST (S-tag-05). The total amount of material spotted on the nitrocellulose membrane is 5 ng/spot. Lane 1: GST-Akt1; Lane 2: GST-Akt2; Lane 3: GST-Akt3; Lane 4: GST-PKAc alpha Monoclonal antibody (Product # MA1-19732); Lane 5: GST-PKAc beta; Lane 6: GST-PKAc gamma; Lane 7: GST-MEK 1; Lane 8: GST.

MA1-19732