Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Other assay [3]
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Validation data
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- Product number
- MA1-19732 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PKA alpha Monoclonal Antibody (6D2.1)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This antibody reacts with human proteinkinase A (an intracellular antigen) catalytic (PKAc) alpha subunit, and weakly with PKAc gamma subunit (both around 40 kDa). The recognized epitope of PKAc alpha is identical between man, sheep, pig, ox and dog.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 6D2.1
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- 4° C, do not freeze
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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.
- 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.
- 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.
- 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.
- 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.