PGP9.5 Mouse Monoclonal Antibody [B1-5-6]
cat.: EM1701-86
Product Type: Mouse monoclonal IgG2a, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Monoclonal
Clone number: B1-5-6
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 25 kDa
Isotype: IgG2a
Immunogen: Synthetic peptide within Human PGP95 aa 174-223 / 223.
Positive control: A-172 cell lysate, SHG-44 cell lysate, U-87 MG cell lysate, SH-SY5Y cell lysate, NCI-H1299 cell lysate, A549 cell lysate, 293T cell lysate, Neuro-2a cell lysate, C6 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, SH-SY-5Y, rat cerebellum tissue, human pancreas tissue, human kidney tissue, mouse brain tissue, Neuro-2a, PC-12.
Subcellular location: Cytoplasm. Endoplasmic reticulum.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:2,000
1:100
1:50-1:200
1:1,000
Uniprot #: SwissProt: P09936 Human | Q9R0P9 Mouse | Q00981 Rat
Alternative names: Epididymis luminal protein 117 Epididymis secretory protein Li 53 HEL 117 HEL S 53 NDGOA Neuron cytoplasmic protein 9.5 OTTHUMP00000218137 OTTHUMP00000218139 OTTHUMP00000218140 OTTHUMP00000218141 Park 5 PARK5 PGP 9.5 PGP9.5 PGP95 Protein gene product 9.5 Ubiquitin C terminal esterase L1 Ubiquitin C terminal hydrolase Ubiquitin C terminal hydrolase L1 Ubiquitin carboxyl terminal esterase L1 Ubiquitin carboxyl terminal hydrolase isozyme L1 Ubiquitin carboxyl-terminal hydrolase isozyme L1 Ubiquitin thioesterase L1 Ubiquitin thiolesterase Ubiquitin thiolesterase L1 UCH-L1 UCHL1 UCHL1_HUMAN
Images
EM1701-86_1.jpg Fig1: Western blot analysis of PGP9.5 on different lysates with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/2,000 dilution.

Lane 1: A-172 cell lysate (20 µg/Lane)
Lane 2: SHG-44 cell lysate (20 µg/Lane)
Lane 3: U-87 MG cell lysate (20 µg/Lane)
Lane 4: SH-SY5Y cell lysate (20 µg/Lane)
Lane 5: NCI-H1299 cell lysate (20 µg/Lane)
Lane 6: A549 cell lysate (20 µg/Lane)
Lane 7: 293T cell lysate (20 µg/Lane)
Lane 8: LNCaP cell lysate (negative) (20 µg/Lane)
Lane 9: K-562 cell lysate (negative) (20 µg/Lane)
Lane 10: Neuro-2a cell lysate (20 µg/Lane)
Lane 11: C6 cell lysate (20 µg/Lane)
Lane 12: PC-12 cell lysate (20 µg/Lane)
Lane 13: Mouse brain tissue lysate (20 µg/Lane)
Lane 14: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 1 minute 50 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 (EM1701-86) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
EM1701-86_2.jpg Fig2: Western blot analysis of PGP9.5 on different lysates with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate (10 µg/Lane)
Lane 2: A549-si PGP9.5 cell lysate (10 µg/Lane)

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 30 seconds; 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 (EM1701-86) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
EM1701-86_3.jpg Fig3: Immunocytochemistry analysis of Neuro-2a cells labeling PGP9.5 with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 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 Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.
EM1701-86_4.jpg Fig4: Immunocytochemistry analysis of PC-12 cells labeling PGP9.5 with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 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 Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.
EM1701-86_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.
EM1701-86_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.
EM1701-86_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.
EM1701-86_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.
EM1701-86_9.jpg Fig9: Flow cytometric analysis of Neuro-2a cells labeling PGP9.5.

Cells were fixed and permeabilized. Then stained with the primary antibody (EM1701-86, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
EM1701-86_10.jpg Fig10: Flow cytometric analysis of PC-12 cells labeling PGP9.5.

Cells were fixed and permeabilized. Then stained with the primary antibody (EM1701-86, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) 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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