Anti-Arc antibody [EPR24093-90] - BSA and Azide free

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cortex (fresh) tissue labeling Arc with ab309193 at 1/50 (10.1 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 on mouse cortex. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309193 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Arc with ab309193 at 1/500 (1.01 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on rat kidney. The section was incubated with ab309193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Arc with ab309193 at 1/500 (1.01 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse kidney. The section was incubated with ab309193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney (fresh) tissue labeling Arc with ab309193 at 1/50 (10.1 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control : confocal image showing no staining on rat kidney (PMID : 12493697). The nuclear counterstain was DAPI (Blue). The section was incubated with ab309193 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh) tissue labeling Arc with ab309193 at 1/50 (10.1 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control : confocal image showing no staining on mouse kidney (PMID : 12493697). The nuclear counterstain was DAPI (Blue). The section was incubated with ab309193 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling Arc with ab309193 at 1/500 (1.01 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse hippocampus. The section was incubated with ab309193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. .Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat hippocampus tissue labeling Arc with ab309193 at 1/500 (1.01 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat hippocampus. The section was incubated with ab309193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh) tissue labeling Arc with ab309193 at 1/50 (10.1 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 on mouse hippocampus. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309193 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody 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-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat cortex primary neuron cells labeling Arc with ab309193 at 1/50 dilution (1ug)/ Right (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). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cortex (fresh) tissue labeling Arc with ab309193 at 1/50 (10.1 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 on rat cortex. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309193 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• WB

Lab

Western blot - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was produced using ab309193, the same antibody clone but in a different buffer.

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

ab181602 was used as a GAPDH loading control at a 1/1000000 dilution.

Negative control : kidney tissue (PMID : 7857651, PMID : 12493697).

All lanes:

Western blot - Anti-Arc antibody [EPR24093-90] (<a href='/en-us/products/primary-antibodies/arc-antibody-epr24093-90-ab309193'>ab309193</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat hippocampus tissue lysate at 20 µg

Lane 6:

Rat kidney 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 1/20000 dilution

Observed band size: 45 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Arc antibody [EPR24093-90] - BSA and Azide free (AB309194)

This data was developed using ab309193, the same antibody clone in a different buffer formulation.

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

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

In Western blot, ab309193 was shown to bind specifically to ARC. Target of interest was observed at 45 kDa in Wild-type Neuro-2a cell lysate (lane 1 and 2) with no signal observed at this size in untreated knockout cell line (lane 3 and 4).

All lanes:

Western blot - Anti-Arc antibody [EPR24093-90] (<a href='/en-us/products/primary-antibodies/arc-antibody-epr24093-90-ab309193'>ab309193</a>) at 1/1000 dilution

Lane 1:

Untreated wild-type Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate at 20 µg

Lane 2:

Wild-type Neuro-2a treated with heat shock at 45 ℃ for 40minutes then recovery for 9 hours, whole cell lysate at 20 µg

Lane 3:

Untreated ARC knockout Neuro-2a, whole cell lysate at 20 µg

Lane 4:

ARC knockout Neuro-2a treated with heat shock at 45 ℃ for 40minutes then recovery for 9 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 1/100000 dilution

Observed band size: 45 kDa,36 kDa

false

Exposure time: 85s