Anti-NPTX2 antibody [EPR24020-38]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling NPTX2 with ab277523 at 1/2000 (0.229 ug/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Positive staining on islet of human pancreas. The section was incubated with ab277523 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling NPTX2 with ab277523 at 1/2000 (0.229 ug/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used. Positive staining on human cerebrum. The section was incubated with ab277523 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

IHC image of NPTX2 staining in a section of frozen normal human pancreas performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab277523, 0.5 ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Negative tissue image : IHC image of NPTX2 staining in a section of frozen normal human heart performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab277523, 0.5 ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

IHC image of NPTX2 staining in a section of frozen normal human cerebral cortex performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab277523, 0.5 ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IP

Supplier Data

Immunoprecipitation - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

NPTX2 was immunoprecipitated from 0.35 mg Human brain tissue lysate with ab277523 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab277523 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human brain tissue lysate 10 ug

Lane 2 : ab277523 IP in Human brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab277523 in human brain tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-NPTX2 antibody [EPR24020-38] (ab277523)

Predicted band size: 47 kDa

Observed band size: 50 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 100% Methanol permeabilized Beta-TC-6 cells labelling NPTX2 with ab277523 at 1/50 (9.16 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Beta-TC-6 cells. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling NPTX2 with ab277523 at 1/2000 (0.229 ug/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Positive staining on mouse cerebrum (PMID : 31772567). The section was incubated with ab277523 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Beta-TC-6 (Mouse pancreas insulinoma beta cell) cells labelling NPTX2 with ab277523 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730)isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling NPTX2 with ab277523 at 1/2000 (0.229 ug/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab277523 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^®RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

NPTX2 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab277523 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab277523 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain tissue lysate 10 ug

Lane 2 : ab277523 IP in Mouse brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab277523 in mouse brain tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-NPTX2 antibody [EPR24020-38] (ab277523)

Predicted band size: 47 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

NPTX2 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 8786423, 28440221).

Exposure time : 15 seconds.

All lanes:

Western blot - Anti-NPTX2 antibody [EPR24020-38] (ab277523) at 1/1000 dilution

All lanes:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 47 kDa

Observed band size: 50-60 kDa

false

• WB

Lab

Western blot - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

NPTX2 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 8786423, 28440221).

Negative : human kidney, human lung (PMID : 8530029, 8786423).

Exposure times : Lane 1 : 15 secondsLane 2-5 : 3 minutes.

All lanes:

Western blot - Anti-NPTX2 antibody [EPR24020-38] (ab277523) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human pancreas tissue lysate at 20 µg

Lane 3:

Human testis tissue lysate at 20 µg

Lane 4:

Human kidney tissue lysate at 20 µg

Lane 5:

Human lung tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 47 kDa

Observed band size: 50-60 kDa

false

• WB

Lab

Western blot - Anti-NPTX2 antibody [EPR24020-38] (AB277523)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

NPTX2 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 8786423, 28440221).

Negative : mouse kidney, mouse lung (PMID : 8530029, 8786423).

Exposure time : 15 seconds.

All lanes:

Western blot - Anti-NPTX2 antibody [EPR24020-38] (ab277523) at 1/1000 dilution

Lane 1:

Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Lane 5:

Mouse lung tissue lysate at 20 µg

Lane 6:

Rat brain tissue lysate at 20 µg

Lane 7:

Rat testis tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 47 kDa

Observed band size: 50-60 kDa

false