Anti-Pentraxin 3/PTX3 antibody [EPR18678-105]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Pentraxin 3/PTX3 antibody [EPR18678-105] (AB190838)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HUVEC (human umbilical vein endothelial cell line) cells labeling Pentraxin 3/PTX3 with ab190838 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluorr^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Pentraxin 3/PTX3 antibody [EPR18678-105] (AB190838)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC (human umbilical vein endothelial cell line) cells labeling Pentraxin 3/PTX3 with ab190838 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HUVEC cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) secondary antibody at 1/200 dilution (red).

-ve control : PBS only, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-Pentraxin 3/PTX3 antibody [EPR18678-105] (AB190838)

Pentraxin 3/PTX3 was immunoprecipitated from 0.35 mg of HUVEC (human umbilical vein endothelial cell line) whole cell lysate with ab190838 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab190838 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : HUVEC whole cell lysate 10 μg (Input).
Lane 2 : ab190838 IP in HUVEC whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab190838 in HUVEC whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.

The molecular weight observed is consistent with what has been described in the literature (PMID : 17389238).

All lanes:

Immunoprecipitation - Anti-Pentraxin 3/PTX3 antibody [EPR18678-105] (ab190838)

Predicted band size: 41 kDa

Observed band size: 42 kDa,47 kDa

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• WB

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Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR18678-105] (AB190838)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR18678-105] (ab190838) at 1/1000 dilution

Lane 1:

Untreated HUVEC (human umbilical vein endothelial cell line) whole cell lysate at 20 µg

Lane 2:

HUVEC treated with 0.5 ug/ml LPS for 4 hours followed by 300 ng/ml BFA for 20 hours, whole cell lysate at 20 µg

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 1/20000 dilution

Predicted band size: 41 kDa

Observed band size: 42 kDa

true

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR18678-105] (AB190838)

Blocking and dilution buffer : 5% NFDM/TBST

Higher concentration of this antibody is required for the detection of Pentraxin 3/PTX3 in untreated cells in WB.

All lanes:

Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR18678-105] (ab190838) at 1/200 dilution

All lanes:

HUVEC (human umbilical vein endothelial cell line) whole cell lysate at 20 µg

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 1/20000 dilution

Predicted band size: 41 kDa

Observed band size: 42 kDa

true

Exposure time: 1min