Name: Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker
SKU: ab108986
Rating: 5 (11 reviews)

Anti-PGP9.5 antibody [EPR4118] ab108986 is a rabbit monoclonal antibody that is used in PGP9.5 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with UCHL1 knockout cell line validation
- Antibody clone EPR4118 is cited in over 120 publications
- One antibody for all your PGP9.5 staining

Related conjugates and formulations

ab196173
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Alexa Fluor® 647 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker
ab300549
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Alexa Fluor® 555 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker
ab302664
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Alexa Fluor® 594 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker
ab302578
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Alexa Fluor® 488 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker
ab305109
Conjugated
Alexa Fluor® 568 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker
ab220823
Carrier free
Anti-PGP9.5 antibody [EPR4118] - BSA and Azide free
ab317591
Conjugated
APC Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker

Images

Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (AB108986), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	IHC-Fr	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Expected	Expected	Tested	Tested
Mouse	Expected	Tested	Tested	Tested	Tested	Tested
Rat	Expected	Tested	Expected	Expected	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/10000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000 - 1/10000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000 - 1/10000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/250	Notes Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

Expected
Expected

Species	Dilution info	Notes
Species Human, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/10000	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Species Mouse	Dilution info 1/100 - 1/10000	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/250 - 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/250 - 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/250 - 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information UCHL1

Function

Deubiquitinase that plays a role in the regulation of several processes such as maintenance of synaptic function, cardiac function, inflammatory response or osteoclastogenesis (PubMed:22212137, PubMed:23359680). Abrogates the ubiquitination of multiple proteins including WWTR1/TAZ, EGFR, HIF1A and beta-site amyloid precursor protein cleaving enzyme 1/BACE1 (PubMed:22212137, PubMed:25615526). In addition, recognizes and hydrolyzes a peptide bond at the C-terminal glycine of ubiquitin to maintain a stable pool of monoubiquitin that is a key requirement for the ubiquitin-proteasome and the autophagy-lysosome pathways (PubMed:12408865, PubMed:8639624, PubMed:9774100). Regulates amyloid precursor protein/APP processing by promoting BACE1 degradation resulting in decreased amyloid beta production (PubMed:22212137). Plays a role in the immune response by regulating the ability of MHC I molecules to reach cross-presentation compartments competent for generating Ag-MHC I complexes (By similarity). Mediates the 'Lys-48'-linked deubiquitination of the transcriptional coactivator WWTR1/TAZ leading to its stabilization and inhibition of osteoclastogenesis (By similarity). Deubiquitinates and stabilizes epidermal growth factor receptor EGFR to prevent its degradation and to activate its downstream mediators (By similarity). Modulates oxidative activity in skeletal muscle by regulating key mitochondrial oxidative proteins (By similarity). Enhances the activity of hypoxia-inducible factor 1-alpha/HIF1A by abrogateing its VHL E3 ligase-mediated ubiquitination and consequently inhibiting its degradation (PubMed:25615526).

Alternative names

Ubiquitin carboxyl-terminal hydrolase isozyme L1, UCH-L1, Neuron cytoplasmic protein 9.5, PGP 9.5, Ubiquitin thioesterase L1, PGP9.5, UCHL1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR4118
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-Fr, IHC-P, IP and WB.

Abcam's high quality manufacturing and validation processes ensure Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986) has been confirmed by Western Blot testing in PGP9.5 knockout Human UCHL1 (PGP9.5) knockout HEK-293T cell lysate ab263773 cells (Human UCHL1 (PGP9.5) knockout HEK-293T cell lysate ab263773).

Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986) has 11 independent reviews from customers.

Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986) specifically detects PGP9.5 (UniProt ID: P09936; Molecular weight: 25kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone EPR4118 - Anti-PGP9.5 antibody [EPR4118] - BSA and Azide free ab220823.

Antibody clone EPR4118 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 647, Alexa Fluor^® 555, Alexa Fluor^® 488, Alexa Fluor^® 594, Alexa Fluor^® 568, APC (Alexa Fluor® 647 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker ab196173, Alexa Fluor® 555 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker ab300549, Alexa Fluor® 488 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker ab302578, Alexa Fluor® 594 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker ab302664, Alexa Fluor® 568 Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker ab305109, APC Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker ab317591).

PGP9.5, also known as UCH-L1/UCHL1, is a crucial protein in neuro research, particularly for its role as a neuronal and neuroendocrine marker. It is widely used to stain neuronal cell bodies and axons in both the central and peripheral nervous systems. Researchers use PGP9.5 to assess nerve fiber density and integrity, making it essential for studying neurodegenerative diseases, small fiber neuropathy and various cancers. Its reliable expression in neurons helps in evaluating neuronal health and pathology in different neurological conditions.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Protein Gene Product 9.5 (PGP9.5) also known as ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) is an enzyme of approximately 27 kDa mass. PGP9.5 is highly expressed in neurons and neuroendocrine cells and is involved in the processing of ubiquitinated proteins. Ubiquitination and deubiquitination are critical for maintaining cellular protein homeostasis. PGP9.5 is present in the central and peripheral nervous systems. Researchers often utilize PGP9.5 as a neuronal marker due to its strong expression in nerve tissues. Antibody clone 13C4 is commonly used in PGP9.5 immunohistochemistry to visualize this protein in histopathology sections.

Biological function summary

PGP9.5 plays a significant role in the ubiquitin-proteasome pathway where it regulates protein degradation. It functions by cleaving ubiquitin from ubiquitin-protein conjugates which can recycle ubiquitin for reuse impacting protein turnover. This process is important for removing misfolded or damaged proteins maintaining cellular integrity. PGP9.5 has not been documented as a part of any large multimeric complex; its action is rather individual yet vital in cellular processes.

Pathways

PGP9.5 is an important participant in the ubiquitin-dependent proteolysis pathway. This pathway controls the degradation of proteins that regulate cell cycle differentiation and neuromodulation. PGP9.5 works in tandem with other proteins such as the proteasome complex to remove proteins tagged for destruction. Another important pathway linked with PGP9.5 is the neuronal development pathway in which its activity supports neuron growth and repair.

Associated diseases and disorders

Defective PGP9.5 function associates mainly with neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. Mutations or dysregulation in PGP9.5 expression are implicated in the pathogenesis of these conditions affecting protein degradation and neuronal health. Other proteins like α-synuclein in Parkinson's disease interact with PGP9.5 pathways indicating their involvement in disease mechanisms. Researchers consider PGP9.5 a potential biomarker for neuronal pathology through methods like PGP9.5 IHC allowing for evaluation of nerve damage and neuroendocrine tumor diagnosis.

Product promise

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19 product images

Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Lanes 1 - 5: Merged signal (red and green). Green - ab108986 observed at 25 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
ab108986 was shown to react with PGP9.5 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line Human UCHL1 (PGP9.5) knockout HEK-293T cell line ab255443 (knockout cell lysate Human UCHL1 (PGP9.5) knockout HEK-293T cell lysate ab263773) was used. Wild-type and PGP9.5 knockout samples were subjected to SDS-PAGE. ab108986 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4^°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986) at 1/1000 dilution
Lane 1: Wild-type HAP1 cell lysate at 20 µg
Lane 2: UCHL1 knockout HAP1 cell lysate at 20 µg
Lane 2: Western blot - Human UCHL1 (PGP9.5) knockout HEK-293T cell line (Human UCHL1 (PGP9.5) knockout HEK-293T cell line ab255443)
Lane 3: SH-SY5Y cell lysate at 20 µg
Lane 4: Wild-type HEK-293T cell lysate at 20 µg
Lane 5: UCHL1 knockout HEK-293T cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 24 kDa
Observed band size: 25 kDa, 37 kDa
Immunoprecipitation - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
PGP9.5 was immunoprecipitated from 0.35 mg Human fetal brain lysate with ab108986 at 1/20 dilution (0.5μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab108986 1/500 dilution (0.17 μg/ml). VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used as the secondary antibody at 1/1000 dilution.
Lane 1: Human fetal brain lysate 10μg
Lane 2: ab108986 IP in Human fetal brain lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab108986 in Human fetal brain lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
All lanes: Immunoprecipitation - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Predicted band size: 24 kDa
Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Lanes 1 - 4: Merged signal (red and green). Green - ab108986 observed at 24 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab108986 was shown to specifically react with UCHL1 in wild-type cells as signal was lost in UCHL1 knockout HAP1 cells. Wild-type and UCHL1 knockout samples were subjected to SDS-PAGE. ab108986 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: UCHL1 knockout HAP1 whole cell lysate at 20 µg
Lane 3: SH-SY5Y whole cell lysate at 20 µg
Lane 4: HEK293 whole cell lysate at 20 µg
Predicted band size: 24 kDa
This image was courtesy of an annonymous customer review
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Fomalin-fixed, paraffin-embedded Dog skin tissue stained for PGP9.5 using ab108986 at 1/500 dilution in immunohistochemical analysis. ImmPRESS™ Anti-Rabbit IgG Polymer Detection Kit was used as the secondary antibody.
Antigen Retrieval: Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Immunohistochemistry (Frozen sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Immunohistochemistry (Frozen sections) analysis of mouse cerebrum tissue sections labeling PGP9.5 with Purified ab108986 at 1/250 (0.5 μg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.
Immunocytochemistry/ Immunofluorescence - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labeling PGP9.5 with ab108986 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Neuro-2a cell line. The nuclear counter stain is DAPI (blue).
The negative control is PBS only.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling PGP9.5 with ab108986, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on human hepatocellular carcinoma. The section was incubated with Anti-DLL3 antibody [EPR22592-18] ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Flow Cytometry (Intracellular) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Y79 (Human retinoblastoma retinoblastoma)cells labelling PGP9.5 with ab108986 at 1/20 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (black)and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluorr^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
All lanes: Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986) at 1/5000 dilution
Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg
Lane 2: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates at 20 µg
Lane 3: C6 (Rat glial tumor glial cell) whole cell lysates at 20 µg
Lanes 4 and 6: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 20 µg
Lane 5: Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates at 20 µg
Lane 7: Mouse brain lysates at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 24 kDa
Observed band size: 25 kDa
Exposure time: 10s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling PGP9.5 with ab108986, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on rat cerebral cortex.The section was incubated with Anti-DLL3 antibody [EPR22592-18] ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling PGP9.5 with ab108986, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on mouse cerebral cortex. The section was incubated with Anti-DLL3 antibody [EPR22592-18] ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
This image is courtesy of an annonymous customer review
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Formalin-fixed, paraffin-embedded Cat lung tissue stained for PGP9.5 using ab108986 at 1/250 dilution in immunohistochemical analysis. The secondary antibody was ImmPRESS™ HRP Universal Antibody (Anti-Mouse IgG/A). Antigen retrieval: Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0.
Flow Cytometry (Intracellular) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Overlay histogram showing SH-SY5Y cells stained with ab108986 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab108986, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr^® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This image is courtesy of a customer review submitted by Carl Hobbs
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Immunohistochemical analysis of rat Jejunum tissue sections labeling PGP9.5 with ab108986 at a dulution of 1/1000. Sections were fixed with Formaldehyde. A Biotin conjugated Goat Anti-Rabbit IgG at 1/300 was used as the secondary antibody. Antigen retrieval was heat mediated using citric acid.
All nerve components of enteric plexuses appear to be very well demonstrated, particularly the fine fibres of the lamina propria and the muscularis mucosa.
Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
All lanes: Western blot - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986) at 1/1000 dilution
Lane 1: Fetal brain cell lysate at 10 µg
Lane 2: Y79 cell lysate at 10 µg
Lane 3: U87-MG cell lysate at 10 µg
Lane 4: SH-SY5Y cell lysate at 10 µg
Lane 5: 293T cell lysate at 10 µg
Secondary
All lanes: HRP labelled goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 24 kDa
Observed band size: 25 kDa
This image is courtesy of a customer review submitted by Carl Hobbs.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
ab108986 staining PGP9.5 in human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with the primary antibody (1/500 in TBS/BSA/azide) for 16 hours at 21°C. A Biotin-conjugated goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.
This image is courtesy of a customer review submitted by Carl Hobbs
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Immunohistochemical analysis of mouse colon tissue sections labeling PGP9.5 with ab108986 at a dulution of 1/1500. Sections were fixed with Formaldehyde. A Biotin conjugated Goat Anti-Rabbit IgG at 1/300 was used as the secondary antibody. Antigen retrieval was heat mediated using citric acid.
All nerve cell/fibre components of enteric plexuses are demonstrated very well.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Immunohistochemical staining of PGP9.5 in paraffin embedded Human glioma tissue, using ab108986 at a 1/250 dilution.
Flow Cytometry (Intracellular) - Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Flow cytometry overlay histogram showing left Neuro2a positive cells and right negative NIH3T3 stained with ab108986 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab108986) (1x 106 in 100μl at 0.2μg/ml (1/10500)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in Neuro2a Fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.