Anti-Agrin antibody [RM2082]

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Agrin antibody [RM2082] (AB324022)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh frozen) tissue labeling Agrin with ab324022 at 1/50 (9.98 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-AGRN (ab324022, green), anti-NeuN (ab190565, white) and anti-GFAP (ab201732, magenta) on mouse cerebrum.
Panel B : anti-AGRN stained in endothelial cells on mouse cerebrum.
Panel C : anti-NeuN stained in neurons of mouse cerebrum.
Panel D : anti-GFAP stained in astrocytes of mouse cerebrum.
The section was incubated in two rounds of staining : in the order of ab324022 and ab190565, ab201732 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Agrin antibody [RM2082] (AB324022)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh frozen) tissue labeling Agrin with ab324022 at 1/50 (9.98 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining in glomeruli of mouse kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab324022 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Agrin antibody [RM2082] (AB324022)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney (fresh frozen) tissue labeling Agrin with ab324022 at 1/50 (9.98 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining in glomeruli of rat kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab324022 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Agrin antibody [RM2082] (AB324022)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung (fresh frozen) tissue labeling Agrin with ab324022 at 1/50 (9.98 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining in endothelial cells on mouse lung. The nuclear counterstain was DAPI (Blue). The section was incubated with ab324022 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Agrin antibody [RM2082] (AB324022)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat lung (fresh frozen) tissue labeling Agrin with ab324022 at 1/50 (9.98 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining in endothelial cells on rat lung. The nuclear counterstain was DAPI (Blue). The section was incubated with ab324022 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Agrin antibody [RM2082] (AB324022)

Agrin was immunoprecipitated from 0.35 mg ab324022 at 1/30 IP in Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab324022 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab324022 at 1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Agrin antibody [RM2082] (ab324022) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 10 µg

Lane 2:

ab324022 at 1/30 IP in Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab324022 in Neuro-2a whole cell 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

Observed band size: 250-300 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Agrin antibody [RM2082] (AB324022)

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

Low expression : RAW 264.7, A20.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18230682, 17586728).

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

All lanes:

Western blot - Anti-Agrin antibody [RM2082] (ab324022) at 1/1000 dilution

Lane 1:

mIMCD3 (mouse inner medullary collecting duct epithelial cell) whole cell lysate at 80 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 80 µg

Lane 3:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 80 µg

Lane 4:

A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate at 80 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

Observed band size: 22 kDa,90 kDa,250-300 kDa,36 kDa

false

Exposure time: 114s

• WB

Supplier Data

Western blot - Anti-Agrin antibody [RM2082] (AB324022)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18230682, 17586728).

The identity of the band at approximately 50 kDa is unknown.

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

All lanes:

Western blot - Anti-Agrin antibody [RM2082] (ab324022) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 80 µg

Lane 2:

Mouse ovary tissue lysate at 80 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

Observed band size: 22 kDa,90 kDa,250-300 kDa,36 kDa

false

Exposure time: 180s