ET1611-54

PP2A alpha + beta Recombinant Rabbit Monoclonal Antibody [SN70-08]

cat.: ET1611-54

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Zebrafish
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	SN70-08

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
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: 36 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human PP2A alpha aa 260-309 / 309.
Positive control:	HeLa cell lysate, A549 cell lysate, A431 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, 293T cell lysate, F9 cell lysate, hybrid fish (crucian-carp) brain tissue lysates, A431, NIH/3T3, PC-12, human kidney tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location:	Cytoplasm, Nucleus, Chromosome, centromere, cytoskeleton, spindle pole.
Recommended Dilutions: WB IF-Cell IHC-P	1:2,000 1:50-1:200 1:1,000
Uniprot #:	SwissProt: P67775 Human \| P62714 Human \| P63330 Mouse \| P62715 Mouse \| P63331 Rat \| P62716 Rat
Alternative names:	PP2A A PP2A alpha PP2A B PP2A beta PP2A-alpha PP2A-beta PP2AA_HUMAN PP2Aalpha PP2AB_HUMAN PP2Abeta PP2Ac PP2CA PP2Calpha PP2CB PPP2CA PPP2CB Protein phosphatase 2 catalytic subunit alpha isoform Protein phosphatase 2 catalytic subunit beta isoform Protein phosphatase type 2A catalytic subunit Replication protein C RP C RP-C RPC Serine/threonine protein phosphatase 2A catalytic subunit alpha isoform Serine/threonine protein phosphatase 2A catalytic subunit beta isoform Serine/threonine-protein phosphatase 2A catalytic subunit alpha isoform

Images

Fig1: Western blot analysis of PP2A alpha + beta on different lysates with Rabbit anti-PP2A alpha + beta antibody (ET1611-54) at 1/2,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: A549 cell lysate (20 µg/Lane)
Lane 3: A431 cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
Lane 5: PC-12 cell lysate (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (20 µg/Lane)
Lane 7: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 6 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 (ET1611-54) 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.

Fig2: Western blot analysis of PP2A alpha + beta on different lysates with Rabbit anti-PP2A alpha + beta antibody (ET1611-54) at 1/2,000 dilution.

Lane 1: A431 cell lysate (10 µg/Lane)
Lane 2: 293T cell lysate (10 µg/Lane)
Lane 3: NIH/3T3 cell lysate (10 µg/Lane)
Lane 4: F9 cell lysate (10 µg/Lane)
Lane 5: PC-12 cell lysate (10 µg/Lane)
Lane 6: Mouse brain tissue lysate (20 µg/Lane)
Lane 7: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 2 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 (ET1611-54) 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: Immunocytochemistry analysis of A431 cells labeling PP2A alpha + beta with Rabbit anti-PP2A alpha + beta antibody (ET1611-54) 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-PP2A alpha + beta antibody (ET1611-54) 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.

Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling PP2A alpha + beta with Rabbit anti-PP2A alpha + beta antibody (ET1611-54) 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-PP2A alpha + beta antibody (ET1611-54) 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.

	Fig5: Immunocytochemistry analysis of PC-12 cells labeling PP2A alpha + beta with Rabbit anti-PP2A alpha + beta antibody (ET1611-54) 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-PP2A alpha + beta antibody (ET1611-54) 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-PP2A alpha + beta antibody (ET1611-54) 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 (ET1611-54) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-PP2A alpha + beta antibody (ET1611-54) 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 (ET1611-54) 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.

Fig8: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-PP2A alpha + beta antibody (ET1611-54) 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 (ET1611-54) 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.