ET1608-30

YAP1 Recombinant Rabbit Monoclonal Antibody [SU33-06]

cat.: ET1608-30

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality:	Monoclonal
Clone number:	SU33-06

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 54 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human YAP1 aa 421-470 / 504.
Positive control:	Hela cell lysate, SiHa cell lysate, HepG2 cell lysate, PC-12 cell lysate, rat liver tissue lysate, HeLa, A549, human colon carcinoma tissue, human breast carcinoma tissue, human kidney tissue, HCT 116 cell lysate, Mouse liver tissue lysate, Mouse kidney tissue lysate, Mouse testis tissue lysate, Mouse skin tissue lysate.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP	1:500-1:2,000 1:50 1:50-1:200 1:200 1:1,000 Use at an assay dependent concentration.
Uniprot #:	SwissProt: P46937 Human \| Q2EJA0 Rat
Alternative names:	65 kDa Yes associated protein 65 kDa Yes-associated protein COB1 YAp 1 YAP 65 YAP YAP1 YAP1_HUMAN YAP2 YAP65 yes -associated protein delta Yes associated protein 1 65kDa Yes associated protein 1 Yes associated protein 2 yes associated protein beta YKI Yorkie homolog

Images

Fig1: Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (ET1608-30) at 1/500 dilution.

Lane 1: Hela cell lysate
Lane 2: SiHa cell lysate
Lane 3: HepG2 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 75 kDa

Exposure time: 2 minutes;

8% 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 (ET1608-30) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (ET1608-30) at 1/1,000 dilution.

Lane 1: PC-12 cell lysate (10 µg/Lane)
Lane 2: Rat liver tissue lysate (20 µg/Lane)

Predicted band size: 54 kDa
Observed band size: 75 kDa

Exposure time: 1 minute;

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 (ET1608-30) 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.

	Fig3: Immunocytochemistry analysis of HeLa cells labeling YAP1 with Rabbit anti-YAP1 antibody (ET1608-30) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-YAP1 antibody (ET1608-30) at 1/50 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 (M1305-2, 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 A549 cells labeling YAP1 with Rabbit anti-YAP1 antibody (ET1608-30) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-YAP1 antibody (ET1608-30) at 1/50 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 (M1305-2, 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: Flow cytometric analysis of HeLa cells labeling YAP1. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1608-30, 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).

	Fig6: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-YAP1 antibody (ET1608-30) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1608-30) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-YAP1 antibody (ET1608-30) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1608-30) 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-YAP1 antibody (ET1608-30) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1608-30) 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.

Fig9: Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (ET1608-30) at 1/1,000 dilution.

Lane 1: HCT 116-si NT cell lysate
Lane 2: HCT 116-si YAP1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 75 kDa

Exposure time: 20 seconds;

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 (ET1608-30) at 1/1,000 dilution was used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig10: Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (ET1608-30) at 1/1,000 dilution.

Lane 1: Mouse liver tissue lysate
Lane 2: Mouse kidney tissue lysate
Lane 3: Mouse testis tissue lysate
Lane 4: Mouse skin tissue lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 70 kDa

Exposure time: 2 minutes; ECL: K1802;

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 (ET1608-30) 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.

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