重组Anti-Kir4.1/KCNJ10抗体[EPR27251-74]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Kir4.1/KCNJ10 with ab312844 at 1/50 (10.14 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 glia cell. 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). Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody was used to counterstain tubulin at 1/200 (2.5ug/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-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling Kir4.1/KCNJ10 with ab312844 at 1/5000 (0.101 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on rat cardiac muscle. The section was incubated with ab312844 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling Kir4.1/KCNJ10 with ab312844 at 1/5000 (0.101 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on mouse cardiac muscle. The section was incubated with ab312844 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Kir4.1/KCNJ10 with ab312844 at 1/5000 (0.101 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on distal tubules in rat kidney (PMID : 8654579). The section was incubated with ab312844 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Kir4.1/KCNJ10 with ab312844 at 1/5000 (0.101 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum (PMID : 11169792). The section was incubated with ab312844 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Kir4.1/KCNJ10 with ab312844 at 1/5000 (0.101 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab312844 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat primary neural/glia cell cells labelling Kir4.1/KCNJ10 with ab312844 at 1/50 dilution (1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left 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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neural/glia cell cells labelling Kir4.1/KCNJ10 with ab312844 at 1/50 dilution (1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left 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.

• WB

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Western blot - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Kir4.1/KCNJ10 is expressed in vivo as a homotetramer (four units of KIR4.1) or a heterotetramer with KIR5.1(two units of KIR4.1 and KIR5.1). (PMID : 33348803; PMID : 15310750 ). This antibody can only detect the tetramer of Kir4.1/KCNJ10. Samples are non-boiled as boiling may cause protein aggregation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Anti-Integrin alpha V (ab179475) control staining at 1/1000 dilution Exposure time : 15 seconds

All lanes:

Western blot - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (ab312844) at 1/1000 dilution

Lane 1:

Mouse brain non-membrane fraction at 20 µg

Lane 2:

Mouse brain membrane fraction at 20 µg

Lane 3:

Rat brain non-membrane fraction at 20 µg

Lane 4:

Rat brain membrane fraction at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 250 kDa

false

Exposure time: 15s

• WB

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Western blot - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : heart (PMID : 11169792). Kir4.1/KCNJ10 is expressed in vivo as a homotetramer (four units of KIR4.1) or a heterotetramer with KIR5.1(two units of KIR4.1 and KIR5.1). (PMID : 33348803; PMID : 15310750 ). This antibody can only detect the tetramer of Kir4.1/KCNJ10. Samples are non-boiled as boiling may cause protein aggregation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 15 seconds

All lanes:

Western blot - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (ab312844) at 1/1000 dilution

Lane 1:

Mouse cerebellum tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Rat cerebellum tissue lysate at 20 µg

Lane 4:

Rat heart tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 250 kDa

false

Exposure time: 15s

• Dot

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Dot Blot - Anti-Kir4.1/KCNJ10 antibody [EPR27251-74] (AB312844)

Dot blot analysis of Kir4.1/KCNJ10 using ab312844 at 1 : 1000 (0.507 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Lane 1 : Kir4.1/KCNJ10 peptide Lane 2 : Kir4.1/KCNJ15 peptide Lane 3 : Kir4.1/KCNJ5 peptide Exposure time : 180 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST