Anti-PCSK9 antibody [RM1106] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PCSK9 antibody [RM1106] - BSA and Azide free (AB315481)

This data was developed using ab315480, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Hepa1-6 (mouse hepatoma epithelial cell) cells labelling PCSK9 with ab315480 at 1/50 (10.32 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in Hepa1-6 cell line.
Low expression : AtT-20.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

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 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PCSK9 antibody [RM1106] - BSA and Azide free (AB315481)

This data was developed using ab315480, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized AtT-20 (mouse pituitary tumor cell, Left) / Hepa1-6 (mouse hepatoma epithelial cell, Right) cells labelling PCSK9 with ab315480 at 1/5000 dilution (0.01 ug)/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 1/5000 dilution was used as the secondary antibody.

Low expression : AtT-20.

• IP

Supplier Data

Immunoprecipitation - Anti-PCSK9 antibody [RM1106] - BSA and Azide free (AB315481)

This data was developed using ab315480, the same antibody clone in a different buffer formulation.

PCSK9 was immunoprecipitated from 0.35 mg Mouse liver tissue lysate with ab315480 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315480 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-PCSK9 antibody [RM1106] (<a href='/en-us/products/primary-antibodies/pcsk9-antibody-rm1106-ab315480'>ab315480</a>) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate

Lane 2:

<a href='/en-us/products/primary-antibodies/pcsk9-antibody-rm1106-ab315480'>ab315480</a> at 1/30 IP in Mouse liver tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/pcsk9-antibody-rm1106-ab315480'>ab315480</a> in mouse liver tissue lysate

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: 23s

• WB

Supplier Data

Western blot - Anti-PCSK9 antibody [RM1106] - BSA and Azide free (AB315481)

This data was developed using ab315480, the same antibody clone in a different buffer formulation.

The expression profile observed is consistent with what has been described in the literature (PMID : 28971102, PMID : 18666258).

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

The identity of the bands between 25 kDa and 35 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PCSK9 antibody [RM1106] (<a href='/en-us/products/primary-antibodies/pcsk9-antibody-rm1106-ab315480'>ab315480</a>) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse ovary tissue lysate at 20 µg

Lane 3:

Mouse heart tissue 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 1/100000 dilution

Observed band size: 62 kDa,36 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-PCSK9 antibody [RM1106] - BSA and Azide free (AB315481)

This data was developed using ab315480, the same antibody clone in a different buffer formulation.

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

The expression profile observed is consistent with what has been described in the literature (PMID : 28971102, PMID : 1819770).

Low expression : AtT20 and Neuro-2a (PMID : 28971102).

Normal serum express low level of PCSK9 (PMID : 18197702).

The identity of the bands between 25 kDa and 35 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PCSK9 antibody [RM1106] (<a href='/en-us/products/primary-antibodies/pcsk9-antibody-rm1106-ab315480'>ab315480</a>) at 1/1000 dilution

Lane 1:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

AtT-20 (mouse corticotroph tumour cell) whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast)whole cell lysate at 20 µg

Lane 4:

Mouse serum tissuse 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 1/100000 dilution

Observed band size: 74 kDa,62 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-PCSK9 antibody [RM1106] - BSA and Azide free (AB315481)

This data was developed using ab315480, the same antibody clone in a different buffer formulation.

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

The expression profile observed is consistent with what has been described in the literature (PMID : 28971102, PMID : 18666258).

This antibody shows lower sensitivity in rat samples than in that of mouse. We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results.

The identity of the bands between 25 kDa and 35 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PCSK9 antibody [RM1106] (<a href='/en-us/products/primary-antibodies/pcsk9-antibody-rm1106-ab315480'>ab315480</a>) at 1/1000 dilution

Lane 1:

Rat liver tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 40 µg

Lane 3:

Rat ovary tissue lysate at 20 µg

Lane 4:

Rat heart tissue 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 1/100000 dilution

Observed band size: 62 kDa

false

Exposure time: 180s