Anti-Glyt2 antibody [EPR25105-74] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Glyt2 with ab303556 at 1/100 (5.08 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on human kidney. The section was incubated with ab303556 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval 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-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human spinal cord tissue labeling Glyt2 with ab303556 at 1/100 (5.08 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human spinal cord. The section was incubated with ab303556 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation. Glyt2 was immunoprecipitated from 0.35 mg Human spinal cord tissue lysate 10 ug with ab303556 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab303556 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Human spinal cord tissue lysate 10 ug Lane 2 : ab303556 IP in Human spinal cord tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab303556 in human spinal cord tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 minutes The lysate of lane 1 is non-boiled as boiling may cause protein aggregates.

All lanes:

Immunoprecipitation - Anti-Glyt2 antibody [EPR25105-74] (<a href='/en-us/products/primary-antibodies/glyt2-antibody-epr25105-74-ab303556'>ab303556</a>) at 1/30 dilution

All lanes:

Human spinal cord 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

Observed band size: 140 kDa

false

Exposure time: 3min

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat brain stem (fresh) tissue labeling Glyt2 with ab303556 at 1/100 (5.08 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 brain stem. The nuclear counterstain was DAPI (Blue). The section was incubated with ab303556 for 60mins 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-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebral cortex (fresh) tissue labeling Glyt2 with ab303556 at 1/100 (5.08 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 cerebral cortex (PMID : 30135938). The nuclear counterstain was DAPI (Blue). The section was incubated with ab303556 for 60mins 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-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebral cortex (fresh) tissue labeling Glyt2 with ab303556 at 1/100 (5.08 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 cerebral cortex (PMID : 30135938). The nuclear counterstain was DAPI (Blue). The section was incubated with ab303556 for 60mins 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-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse brain stem (fresh) tissue labeling Glyt2 with ab303556 at 1/100 (5.08 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 brain stem. The nuclear counterstain was DAPI (Blue). The section was incubated with ab303556 for 60mins 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-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse brainstem tissue labeling Glyt2 with ab303556 at 1/100 (5.08 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse brainstem. The section was incubated with ab303556 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval 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-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling Glyt2 with ab303556 at 1/100 (5.08 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse cerebellum. The section was incubated with ab303556 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation. Glyt2 was immunoprecipitated from 0.35 mg Mouse spinal cord tissue lysate 10 ug with ab303556 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab303556 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse spinal cord tissue lysate 10 ug Lane 2 : ab303556 IP in Mouse spinal cord tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab303556 in mouse spinal cord tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 minutes The lysate of lane 1 is non-boiled as boiling may cause protein aggregates.

All lanes:

Immunoprecipitation - Anti-Glyt2 antibody [EPR25105-74] (<a href='/en-us/products/primary-antibodies/glyt2-antibody-epr25105-74-ab303556'>ab303556</a>) at 1/1000 dilution

All lanes:

Mouse spinal cord 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

Observed band size: 100 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Glyt2 antibody [EPR25105-74] - BSA and Azide free (AB303557)

This data was developed using ab303556, the same antibody clone in a different buffer formulation. Blocking and dilution buffer and concentration : 5% NFDM/TBST.  The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 7751957; PMID : 30135938). Negative control : kidney (PMID : 7751957) Low expression : hippocampus (PMID : 7751957) Samples are non-boiled as boiling may cause protein aggregates.

All lanes:

Western blot - Anti-Glyt2 antibody [EPR25105-74] (<a href='/en-us/products/primary-antibodies/glyt2-antibody-epr25105-74-ab303556'>ab303556</a>) at 1/1000 dilution

Lane 1:

Human spinal cord tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Lane 4:

Mouse brain stem tissue lysate at 20 µg

Lane 5:

Mouse hippocampus tissue lysatE at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 87 kDa

Observed band size: 100 kDa

false

Exposure time: 15s