Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

Composite multiplex immunofluorescence staining of APP, PSD95 and Tau staining in a section of formalin-fixed paraffin-embedded human Alzheimer’s brain*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with EDTA (Ph9.0) using retrieval settings of 100°C for 40 minutes. The section was then incubated at room temperature for 1 hour with ab201060 at 1µg/ml dilution (shown in green), ab317793 at 1µg/ml (shown in magenta), and ab317841 at 1µg/ml (shown in yellow). Then incubated for 1 hour with ab150118 Goat Anti-Mouse IgG H&L (Alexa Fluor® 555) preadsorbed 1/1000, ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 1/1000, and ab150135 Donkey Anti-Goat IgG H&L (Alexa Fluor® 647) preadsorbed 1/1000. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium ^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

Immunofluorescence staining of PSD95 in sections of formalin-fixed paraffin-embedded rat brain (positive) and rat liver (negative). Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab317803 at 1/100 dilution, and then incubated for 1 hour with ab150135 Donkey Anti-Goat IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 dilution (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab317793).

Immunohistochemical analysis of 4% PFA-fixed frozen Mouse liver (perfused fixed) tissue labeling PSD95 with ab317793 at 1/200 dilution followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green).

Negative control : confocal image showing no staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317793 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 ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab317793).

Immunohistochemical analysis of 4% PFA-fixed frozen Rat liver (perfused fixed) tissue labeling PSD95 with ab317793 at 1/200 dilution followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green).

Negative control : confocal image showing no staining on rat liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317793 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 ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

Immunofluorescence staining of PSD95 in sections of formalin-fixed paraffin-embedded mouse brain (positive) and mouse liver (negative). Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab317803 at 1/100 dilution, and then incubated for 1 hour with ab150135 Donkey Anti-Goat IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 dilution (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab317793).

Immunohistochemical analysis of 4% PFA-fixed frozen Mouse retina (perfused fixed) tissue labeling PSD95 with ab317793 at 1/200 dilution followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green).

Confocal image showing positive staining on mouse retina tissue. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317793 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 ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab317793).

Immunohistochemical analysis of 4% PFA-fixed frozen Rat retina (perfused fixed) tissue labeling PSD95 with ab317793 at 1/200 dilution followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green).

Confocal image showing positive staining on rat retina tissue. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317793 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 ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IP

Lab

Immunoprecipitation - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab317793).

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

In Western blot, ab317793 was used at 1/1000 dilution.

All lanes:

Immunoprecipitation - Anti-PSD95 antibody [EPR23124-118] - Synaptic Marker – Goat IgG (Chimeric) (<a href='/ko/products/primary-antibodies/psd95-antibody-epr23124-118-synaptic-marker-goat-igg-chimeric-ab317793'>ab317793</a>) at 1/30 dilution

Lane 1:

Rat brain lysate (Input) at 10 µg

Lane 2:

Rat brain lysate (+)

Lane 3:

Goat polyclonal IgG (<a href='/ko/products/primary-antibodies/goat-igg-polyclonal-isotype-control-ab37373'>ab37373</a>) instead of <a href='/ko/products/primary-antibodies/psd95-antibody-epr23124-118-synaptic-marker-goat-igg-chimeric-ab317793'>ab317793</a> in rat brain tissue lysate

Secondary

All lanes:

Peroxidase-Conjugated Rabbit anti-Goat IgG (H+L) at 1/1000 dilution

false

Exposure time: 3s

• IP

Lab

Immunoprecipitation - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab317793).

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

In Western blot, ab317793 was used at 1/1000 dilution.

All lanes:

Immunoprecipitation - Anti-PSD95 antibody [EPR23124-118] - Synaptic Marker – Goat IgG (Chimeric) (<a href='/ko/products/primary-antibodies/psd95-antibody-epr23124-118-synaptic-marker-goat-igg-chimeric-ab317793'>ab317793</a>) at 1/30 dilution

Lane 1:

Mouse brain lysate (Input) at 10 µg

Lane 2:

Mouse brain lysate (+)

Lane 3:

Goat polyclonal IgG (<a href='/ko/products/primary-antibodies/goat-igg-polyclonal-isotype-control-ab37373'>ab37373</a>) instead of <a href='/ko/products/primary-antibodies/psd95-antibody-epr23124-118-synaptic-marker-goat-igg-chimeric-ab317793'>ab317793</a> in mouse brain tissue lysate

Secondary

All lanes:

Peroxidase-Conjugated Rabbit anti-Goat IgG (H+L) at 1/1000 dilution

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab317793).

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) was used (36KDa).

Negative control : liver, testis and spleen.

All lanes:

Western blot - Anti-PSD95 antibody [EPR23124-118] - Synaptic Marker – Goat IgG (Chimeric) (<a href='/ko/products/primary-antibodies/psd95-antibody-epr23124-118-synaptic-marker-goat-igg-chimeric-ab317793'>ab317793</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human testis tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Rabbit anti-Goat IgG (H+L) at 1/5000 dilution

Observed band size: 100 kDa,75 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-PSD95 antibody [EPR23124-118] – Goat IgG (Chimeric) – BSA and Azide Free (AB317803)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab317793).

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) was used (36KDa).

Negative control : liver.

All lanes:

Western blot - Anti-PSD95 antibody [EPR23124-118] - Synaptic Marker – Goat IgG (Chimeric) (<a href='/ko/products/primary-antibodies/psd95-antibody-epr23124-118-synaptic-marker-goat-igg-chimeric-ab317793'>ab317793</a>) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 20 µg

Lane 2:

Mouse liver lysate at 20 µg

Lane 3:

Rat brain lysate at 20 µg

Lane 4:

Rat liver lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Rabbit anti-Goat IgG (H+L) at 1/5000 dilution

Observed band size: 100 kDa,75 kDa

false