Anti-NP-I antibody [EPR25683-17] (BSA and Azide free)

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-NP-I antibody [EPR25683-17] (BSA and Azide free) (AB300416)

This data was developed using ab300404, the same antibody clone in a different buffer formulation.Flow cytometric analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labelling NPTX1 with ab300404 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NP-I antibody [EPR25683-17] (BSA and Azide free) (AB300416)

This data was developed using ab300404, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized IMR-32 (human neuroblastoma neuroblast) cells labelling NPTX1 with ab300404 at 1/50 (9.56 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasm staining in IMR-32 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IP

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Immunoprecipitation - Anti-NP-I antibody [EPR25683-17] (BSA and Azide free) (AB300416)

This data was developed using ab300404, the same antibody clone in a different buffer formulation. NPTX1 was immunoprecipitated from 0.35 mg Human hypothalamus tissue lysate 10 ug with ab300404 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300404 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Human hypothalamus tissue lysate 10 ug

Lane 2 : ab300404 IP in Human hypothalamus tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300404 in Human hypothalamus tissue lysate

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

Exposure time : 3.25 seconds

All lanes:

Immunoprecipitation - Anti-NP-I antibody [EPR25683-17] (<a href='/en-us/products/primary-antibodies/np-i-antibody-epr25683-17-ab300404'>ab300404</a>) at 1/1000 dilution

All lanes:

Human hypothalamus tissue lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 3.25s

• WB

Supplier Data

Western blot - Anti-NP-I antibody [EPR25683-17] (BSA and Azide free) (AB300416)

This data was developed using ab300404, the same antibody clone in a different buffer formulation. Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-NP-I antibody [EPR25683-17] (<a href='/en-us/products/primary-antibodies/np-i-antibody-epr25683-17-ab300404'>ab300404</a>) at 1/1000 dilution

Lane 1:

Hela (Human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control, whole cell lysate

Lane 2:

Hela transfected with siRNA specifically targeti NPTX1, whole cell lysate 20

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 50 kDa

false

Exposure time: 169s

• WB

Supplier Data

Western blot - Anti-NP-I antibody [EPR25683-17] (BSA and Azide free) (AB300416)

This data was developed using ab300404, the same antibody clone in a different buffer formulation. Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-NP-I antibody [EPR25683-17] (<a href='/en-us/products/primary-antibodies/np-i-antibody-epr25683-17-ab300404'>ab300404</a>) at 1/1000 dilution

Lane 1:

Human hypothalamus tissue lysate

Lane 2:

Human heart tissue lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 50 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-NP-I antibody [EPR25683-17] (BSA and Azide free) (AB300416)

This data was developed using ab300404, the same antibody clone in a different buffer formulation. Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-NP-I antibody [EPR25683-17] (<a href='/en-us/products/primary-antibodies/np-i-antibody-epr25683-17-ab300404'>ab300404</a>) at 1/1000 dilution

All lanes:

IMR-32 (Human neuroblastoma neuroblast) whole cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 50 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-NP-I antibody [EPR25683-17] (BSA and Azide free) (AB300416)

This data was developed using ab300404, the same antibody clone in a different buffer formulation. Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-NP-I antibody [EPR25683-17] (<a href='/en-us/products/primary-antibodies/np-i-antibody-epr25683-17-ab300404'>ab300404</a>) at 1/1000 dilution

Lane 1:

His-tagged human NPTX1 recombinant protein

Lane 2:

His-tagged human NPTX2 recombinant protein

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 50 kDa

false

Exposure time: 3.25s