Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free
See all NPTX2 primary antibodies
Description

Rabbit monoclonal [EPR24020-38] to NPTX2 - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
IHC-P	Mouse
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human brain, pancreas and testis tissue lysates; SH-SY5Y whole cell lysate; Mouse brain, kidney and testis tissue lysates; Rat brain and testis tissue lysates. IHC-P: Human pancreas and cerebrum tissue; Mouse cerebrum tissue; Rat cerebrum tissue. ICC: Beta-TC-6 cells. Flow cyt: Beta-TC-6 cells. IP: Human brain tissue lysate; Mouse brain tissue lysate.
General notes
ab277533 is the carrier-free version of ab277523. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab277533 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C.
Storage buffer

Constituent: 100% PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24020-38
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Immunocytochemistry - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Western blot - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

All lanes : Anti-NPTX2 antibody [EPR24020-38] (ab277523) at 1/1000 dilution

Lane 1 : Beta-TC-6 (mouse pancreas insinoma beta cell) whole cell lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Mouse testis tissue lysate
Lane 4 : Mouse kidney tissue lysate
Lane 5 : Mouse lu tissue lysate 20
Lane 6 : Rat brain tissue lysate
Lane 7 : Rat testis tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

Predicted band size: 47 kDa
Observed band size: 50-60 kDa
why is the actual band size different from the predicted?

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Immunoprecipitation - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Flow Cytometry - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Western blot - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

All lanes : Anti-NPTX2 antibody [EPR24020-38] (ab277523) at 1/1000 dilution

Lane 1 : Human brain tissue lysate
Lane 2 : Human pancreas tissue lysate
Lane 3 : Human testis tissue lysate
Lane 4 : Human kidney tissue lysate
Lane 5 : Human lung tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 47 kDa
Observed band size: 50-60 kDa why is the actual band size different from the predicted?

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Western blot - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

Anti-NPTX2 antibody [EPR24020-38] (ab277523) at 1/1000 dilution + SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

Predicted band size: 47 kDa
Observed band size: 50-60 kDa why is the actual band size different from the predicted?

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Immunoprecipitation - Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)

This data was developed using ab277523, the same antibody clone in a different buffer formulation.

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.
Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)