ET7109-92

PKA R2 Recombinant Rabbit Monoclonal Antibody [JE49-49]

cat.: ET7109-92

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	JE49-49

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 46 kDa.
Isotype:	IgG
Immunogen:	Recombinant protein within Human PKA R2 aa 272-404 / 404.
Positive control:	A549 cell lysate, A549 cell lysate, HeLa cell lysate, MCF7 cell lysate, K-562 cell lysate, PC-12 cell lysate, human colon cancer tissue, human kidney tissue, human liver cancer tissue, human skin tissue, human breast tissue, human placenta tissue, PC-12.
Subcellular location:	Cytoplasm, Cell membrane.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:2,000 1:50-1:200 1:100 1:1,000
Uniprot #:	SwissProt: P13861 Human \| P12368 Rat
Alternative names:	Alternative names cAMP dependent protein kinase regulatory subunit alpha 2 cAMP dependent protein kinase regulatory subunit RII alpha cAMP dependent protein kinase type II alpha regulatory chain cAMP dependent protein kinase type II alpha regulatory subunit cAMP-dependent protein kinase type II-alpha regulatory subunit KAP2 KAP2_HUMAN MGC3606 PKR 2 PKR2 PRKA R2 PRKAR 2 PRKAR2 PRKAR2A Protein kinase A RII alpha subunit Protein kinase cAMP dependent regulatory type II alpha

Images

Fig1: Western blot analysis of PKA R2 on different lysates with Rabbit anti-PKA R2 antibody (ET7109-92) at 1/1,000 dilution.

Lane 1: A549-WT cell lysate
Lane 2: A549-KD PKA R2 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 46 kDa
Observed band size: 50 kDa

Exposure time: 30 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7109-92) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of PKA R2 on different lysates with Rabbit anti-PKA R2 antibody (ET7109-92) at 1/2,000 dilution.

Lane 1: A549 cell lysate
Lane 2: HeLa cell lysate
Lane 3: MCF7 cell lysate
Lane 4: K-562 cell lysate
Lane 5: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 46 kDa
Observed band size: 50 kDa

Exposure time: 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7109-92) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-PKA R2 antibody (ET7109-92) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7109-92) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig4: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-PKA R2 antibody (ET7109-92) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7109-92) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded human liver cancer tissue using anti-PKA R2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7109-92, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig6: Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-PKA R2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7109-92, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig7: Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-PKA R2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7109-92, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig8: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-PKA R2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7109-92, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig9: Immunocytochemistry analysis of PC-12 cells labeling PKA R2 with Rabbit anti-PKA R2 antibody (ET7109-92) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PKA R2 antibody (ET7109-92) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig10: Flow cytometric analysis of PC-12 cells labeling PKA R2. Cells were fixed and permeabilized. Then stained with the primary antibody (ET7109-92, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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