ET1611-45

PARK7/DJ1 Recombinant Rabbit Monoclonal Antibody [SN07-21]

cat.: ET1611-45

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:	SN07-21

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: 20 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human PARK7 aa 11-60 / 189.
Positive control:	HeLa cell lysate, SH-SY5Y cell lysate, HepG2 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, HeLa, human breast tissue, human kidney tissue, human pancreas tissue, mouse prostate tissue, mouse pancreas tissue.
Subcellular location:	Cell membrane, Cytoplasm, Endoplasmic reticulum, Membrane, Mitochondrion, Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP	1:1,000-1:2,000 1:100 1:100-1:500 1:100-1:500 1:50-1:100 Use at an assay dependent concentration.
Uniprot #:	SwissProt: Q99497 Human \| Q99LX0 Mouse \| O88767 Rat
Alternative names:	CAP1 DJ-1 DJ1 DJ1 protein Epididymis secretory sperm binding protein Li 67p FLJ27376 FLJ34360 FLJ92274 HEL S 67p Oncogene DJ1 OTTHUMP00000001348 OTTHUMP00000001349 OTTHUMP00000001350 OTTHUMP00000001351 PARK7 PARK7_HUMAN Parkinson disease (autosomal recessive, early onset) 7 Parkinson disease protein 7 Parkinson protein 7 Protein DJ-1 SP22

Images

Fig1: Western blot analysis of PARK7/DJ1 on different lysates with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: SH-SY5Y cell lysate
Lane 3: HepG2 cell lysate
Lane 4: Mouse brain tissue lysate
Lane 5: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 20 kDa
Observed band size: 20 kDa

Exposure time: 20 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 (ET1611-45) 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 PARK7/DJ1 on different lysates with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/1,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si PARK7/DJ1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 20 kDa
Observed band size: 20 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 (ET1611-45) 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 PARK7/DJ1 with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/100 dilution. Cells were fixed in 100% precooled methanol 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-PARK7/DJ1 antibody (ET1611-45) 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 (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: Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-PARK7/DJ1 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 (ET1611-45, 1/50) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PARK7/DJ1 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 (ET1611-45, 1/50) 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 pancreas tissue using anti-PARK7/DJ1 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 (ET1611-45, 1/50) 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 mouse prostate tissue using anti-PARK7/DJ1 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 (ET1611-45, 1/50) 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 mouse pancreas tissue using anti-PARK7/DJ1 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 (ET1611-45, 1/50) 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: Flow cytometric analysis of PARK7/DJ1 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1611-45, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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