Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [2]
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Validation data
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- Product number
- PA5-21842 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PKA alpha Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: HeLaS3, NIH-3T3, C2C12. Predicted reactivity: Mouse (98%), Rat (98%), Xenopus laevis (94%), Dog (99%), Rabbit (97%), Sheep (99%), Bovine (99%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using PKA alpha Polyclonal Antibody (Product # PA5-21842). Sample (30 µg of whole cell lysate). A: HeLa S3. 12% SDS PAGE. PKA alpha Polyclonal Antibody (Product # PA5-21842) diluted at 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using PKA alpha Polyclonal Antibody (Product # PA5-21842). Sample (30 µg of whole cell lysate). Lane A: NIH-3T3. Lane B: C2C12. 10% SDS PAGE. PKA alpha Polyclonal Antibody (Product # PA5-21842) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of PRKACA was achieved by transfecting NIH: OVCAR-3 with PRKACA specific siRNAs (Silencer® select Product # s8587, s8586). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the PRKACA knockdown cells (Lane 1), non-specific scrambled siRNA transfected cells (Lane 1) and untransfected cells (Lane 3). The blot was probed with PKA alpha Polyclonal Antibody (Product # PA5-21842, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown i histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to PRKACA. An uncharacterized band was observed at ~60 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using anti-PKA alpha Polyclonal Antibody (Product # PA5-21842) and a 40 kDa band corresponding to PRKACA was observed across cell lines tested along with an uncharacterized band at ~70kDa. Whole cell extracts (30 µg lysate) of NIH: OVCAR-3 (Lane 1), A549 (Lane 2) and HeLa (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 (1:500 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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
- Immunofluorescent analysis of PRKACA in paraformaldehyde-fixed HeLa cells using a PRKACA polyclonal antibody (Product # PA5-21842) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Confocal immunofluorescence analysis (Olympus FV10i) of paraformaldehyde-fixed HeLa, using PKA alpha antibody (Product # PA5-21842) (Green) at 1:500 dilution. Alpha-tubulin filaments were labeled with (Product # MA1-25054) (Red) at 1:2,000.