Anti-NTT4 antibody [EPR28520-13]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling NTT4 with ab313747 at 1/500 (1.09 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebrum. The section was incubated with ab313747 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling NTT4 with ab313747 at 1/500 (1.09 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : no staining on human liver. The section was incubated with ab313747 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling NTT4 with ab313747 at 1/500 (1.09 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : no staining on human kidney. The section was incubated with ab313747 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-NTT4 antibody [EPR28520-13] (AB313747)

NTT4 was immunoprecipitated from 0.35 mg Human cerebellum tissue lysate with ab313747 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313747 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Human cerebellum tissue lysate 10 ug Lane 2 : Human cerebellum tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313747 in human cerebellum tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-NTT4 antibody [EPR28520-13] (ab313747) at 1/30 dilution

All lanes:

Human cerebellum 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: 8s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling NTT4 with ab313747 at 1/1000 (0.545 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebellum. The section was incubated with ab313747 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling NTT4 with ab313747 at 1/1000 (0.545 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cerebellum. The section was incubated with ab313747 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling NTT4 with ab313747 at 1/1000 (0.545 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum. The section was incubated with ab313747 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh) tissue labeling NTT4 with ab313747 at 1/50 (10.9 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 cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab313747 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). The nuclear counterstain was DAPI (Blue). 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-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum (fresh) tissue labeling NTT4 with ab313747 at 1/50 (10.9 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 cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab313747 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). 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling NTT4 with ab313747 at 1/50 (10.9 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 positive staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse kidney (fresh) tissue labeling NTT4 with ab313747 at 1/50 (10.9 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 : 23672601). The nuclear counterstain was DAPI (Blue). The section was incubated with ab313747 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). 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-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney (fresh) tissue labeling NTT4 with ab313747 at 1/50 (10.9 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 : 23672601). The nuclear counterstain was DAPI (Blue). The section was incubated with ab313747 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). 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-NTT4 antibody [EPR28520-13] (AB313747)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling NTT4 with ab313747 at 1/1000 (0.545 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cerebrum. The section was incubated with ab313747 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-NTT4 antibody [EPR28520-13] (AB313747)

NTT4 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab313747 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313747 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse brain tissue lysate 10 ug Lane 2 : Mouse brain tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313747 in mouse brain tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-NTT4 antibody [EPR28520-13] (ab313747) at 1/30 dilution

All lanes:

Mouse brain 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: 8s

• WB

Supplier Data

Western blot - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : liver, kidney (PMID : 23672601). The expression profile observed is consistent with what has been described in literature (PMID : 23672601). The identity of the lower MW band at approximately 25 kDa is unknown. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Samples are non-boiled as boiling may cause protein aggregation. Exposure time : Lane1-2 : 15 seconds; Lane 3-5 : 26 seconds

All lanes:

Western blot - Anti-NTT4 antibody [EPR28520-13] (ab313747) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human hypothalamus tissue lysate at 20 µg

Lane 3:

Human spinal cord tissue lysate at 20 µg

Lane 4:

Human kidney tissue lysate at 20 µg

Lane 5:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 75 kDa

false

• WB

Supplier Data

Western blot - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Blocking and diluting buffer and concentration : 5% NFDM/TBST NTT4 detected a shift from 75kDa to 65kDa band after deglycosylation treatment. In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution. Exposure time : 15 seconds

All lanes:

Western blot - Anti-NTT4 antibody [EPR28520-13] (ab313747) at 1/1000 dilution

Lane 1:

Untreated mouse brain tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate treated with Protein treated with Peptide:N-glycosidase F (PNGase F) 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: 75 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-NTT4 antibody [EPR28520-13] (AB313747)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : liver, kidney (PMID : 23672601). The expression profile observed is consistent with what has been described in literature (PMID : 23672601). The identity of the lower MW band at approximately 25 kDa is unknown. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Samples are non-boiled as boiling may cause protein aggregation. Exposure time : 26 seconds

All lanes:

Western blot - Anti-NTT4 antibody [EPR28520-13] (ab313747) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse cerebellum tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat kidney tissue lysate at 20 µg

Lane 7:

Mouse hippocampus tissue lysate at 20 µg

Lane 8:

Mouse spinal cord tissue lysate at 20 µg

Lane 9:

Rat cerebellum 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: 75 kDa

false

Exposure time: 26s