- PA5-22129 - Invitrogen Antibodies PEX19 antibody

Submitted references Dual role of USP30 in controlling basal pexophagy and mitophagy.

Marcassa E, Kallinos A, Jardine J, Rusilowicz-Jones EV, Martinez A, Kuehl S, Islinger M, Clague MJ, Urbé S
EMBO reports 2018 Jul;19(7)

The peroxisome biogenesis factors posttranslationally target reticulon homology domain-containing proteins to the endoplasmic reticulum membrane.

Yamamoto Y, Sakisaka T
Scientific reports 2018 Feb 2;8(1):2322

Parkinson's disease-associated mutant VPS35 causes mitochondrial dysfunction by recycling DLP1 complexes.

Wang W, Wang X, Fujioka H, Hoppel C, Whone AL, Caldwell MA, Cullen PJ, Liu J, Zhu X
Nature medicine 2016 Jan;22(1):54-63

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of PEX19 using 30 µg of A) Raji (B) K562 (C) THP-1 and D) NCI-H929 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a PEX19 polyclonal antibody (Product # PA5-22129) at a dilution of 1:10,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of PEX19 was performed by separating 30 µg of various whole cell extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a PEX19 Polyclonal Antibody (Product # PA5-22129) at a dilution of 1:10000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PEX19 Polyclonal Antibody detects PEX19 protein by western blot analysis. A. 30 µg PC-12 whole cell lysate/extract. B. 30 µg Rat-2 whole cell lysate/extract.10% SDS-PAGE. PEX19 Polyclonal Antibody (Product # PA5-22129) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of PEX19 was performed by separating 30 µg of various whole cell extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a PEX19 Polyclonal Antibody (Product # PA5-22129) at a dilution of 1:10000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PEX19 Polyclonal Antibody (Product # PA5-22129). Sample (50 µg of whole cell lysate). Lane A: mouse liver. 12% SDS PAGE. PEX19 Polyclonal Antibody (Product # PA5-22129) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PEX19 Polyclonal Antibody (Product # PA5-22129). Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with PEX19 Polyclonal Antibody (Product # PA5-22129) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PEX19 Polyclonal Antibody detects PEX19 protein by western blot analysis. A. 30 µg PC-12 whole cell lysate/extract. B. 30 µg Rat-2 whole cell lysate/extract.10% SDS-PAGE. PEX19 Polyclonal Antibody (Product # PA5-22129) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of PEX19 was performed by separating 30 µg of various whole cell extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a PEX19 Polyclonal Antibody (Product # PA5-22129) at a dilution of 1:10000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of PEX19 was achieved by transfecting HeLa cells with PEX19 specific siRNAs (Silencer® select Product # s11614, s11613). Western blot analysis (Fig. a) was performed using whole cell extracts from the PEX19 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PEX19 Polyclonal Antibody (Product # PA5-22129, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to PEX19.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), PC-12 (Lane 2), Raji (Lane 3), K-562 (Lane 4), Jurkat (Lane 5), THP-1 (Lane 6), NTERA-2 (Lane 7) and LNCap (Lane 8). The blot was probed with Anti- PEX19 Polyclonal Antibody (Product # PA5-22129, 1µg/ml) and detected by chemiluminescence using Goat anti Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 37 kDa band corresponding to PEX19 was detected across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of PEX19 in methanol-fixed HeLa cells using a PEX19 polyclonal antibody (Product # PA5-22129) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Confocal immunofluorescence analysis (Olympus FV10i) of methanol-fixed HeLa, using PEX19 antibody (Product # PA5-22129) (Green) at 1:500 dilution. Alpha-tubulin filaments were labeled with (Product # MA1-25054) (Red) at 1:2,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of PEX19 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PEX19 Rabbit Polyclonal Antibody (Product # PA5-22129) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PEX19 Polyclonal Antibody detects PEX19 protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded mouse kidney. PEX19 stained by PEX19 Polyclonal Antibody (Product # PA5-22129) diluted at 1:1,000. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PEX19 Polyclonal Antibody detects PEX19 protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded rat liver. PEX19 stained by PEX19 Polyclonal Antibody (Product # PA5-22129) diluted at 1:1,000. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure EV3 Basal autophagic flux and peroxisome abundance are not affected in USP30 KO cells Representative images of hTERT-RPE1 cells transfected with Keima-SKL and CFP-LAMP1 for 48 h, fixed and analysed by 3i-spinning disk confocal microscopy. Scale bar: 10 mum. Arrows indicate Keima-SKL ""red"" puncta that colocalise with the lysosomal marker CFP-LAMP1. Representative Western blot of cells analysed in Fig 5 B. Keima-SKL ""red"" puncta in USP30 KO or WT hTERT-RPE1 cells. Keima-SKL was either transfected on its own or together with USP30-GFP and USP30C77S-GFP for 48 h. Two independent experiments were analysed, 20 cells per experiment, mean +- range. Quantification of the percentage of mt-Keima ""red"" puncta in USP30 KO6 or WT1 hTERT-RPE1 cells ( n = 2 independent experiments, 20 cells per experiment, mean +- range). hTERT-RPE1 USP30 WT and KO cells (WT1, WT3, KO2 and KO6) were transfected with RFP-GFP-LC3b for 48 h prior to imaging by 3i-spinning disk confocal microscopy. Shown are representative images and the numbers of RFP-GFP-LC3b puncta per cell as an assessment of basal autophagic flux. Two independent experiments were analysed, 20 cells per experiment, mean +- range. hTERT-RPE1 cells were transfected with non-targeting (NT1) or USP30 targeting siRNA (D1 and D3) for 72 h and lysed in RIPA buffer. Protein samples were analysed by SDS-PAGE and Western blots probed for catalase, PMP70, PEX5 and PEX19. hTERT-RPE1 USP30 WT and KO cells (WT1, WT3, KO2 and KO6) were lysed i

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Identification of PEX19 as a protein that selectively binds to the nascent RHD-containing proteins. ( A ) Purification of Arl6IP1-binding proteins from the rabbit reticulocyte lysates. The rabbit reticulocyte lysate synthesizing FLAG-Arl6IP1 was incubated with the anti-FLAG mAb-immobilized agarose. To check the contamination, the rabbit reticulocyte lysate without synthesis of FLAG-Arl6IP1 (control lysate) was also incubated with the anti-FLAG mAb-immobilized agarose. The bound proteins were eluted by competition with a large excess of free FLAG peptide and subjected to SDS-PAGE followed by silver staining. p120, p60, p55, p40 and p29 were identified as Bat3 (also known as Bag6), proteasome 26 S subunit ATPase1, SRP54, PEX19 and ribosomal protein L10a, respectively, by mass spectrometry. ( B ) Selective binding of PEX19 to the nascent RHD-containing proteins. The rabbit reticulocyte lysates synthesizing FLAG-Arl6IP1, FLAG-Rtn4C, FLAG-atlastin-1 and FLAG-syntaxin-1 were supplemented with 1 mM puromycin, followed by incubation with anti-FLAG mAb-immobilized agarose. The bound proteins were eluted with the SDS sample buffer with boiling and subjected to SDS-PAGE followed by Western blotting with the anti-PEX19 pAb and the anti-FLAG pAb.

PA5-22129

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