Anti-LMX1b antibody [EPR24053-161]

• IP

Supplier Data

Immunoprecipitation - Anti-LMX1b antibody [EPR24053-161] (AB259926)

LMX1b was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney) transfected with mouse LMX1b expression vector containing a myc-His-tag®, whole cell lysate 10 ug with ab259926 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259926 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HEK-293T (human embryonic kidney) transfected with mouse LMX1b expression vector containing a myc-His-tag®, whole cell lysate 10 ug

Lane 2 : ab259926 IP in HEK-293T transfected with mouse LMX1b expression vector containing a myc-His-tag®, whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab259926 in HEK-293T transfected with mouse LMX1b expression vector containing a myc-His-tag®, whole cell lysate

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

Exposure time : 32 seconds

All lanes:

Immunoprecipitation - Anti-LMX1b antibody [EPR24053-161] (ab259926)

Predicted band size: 42 kDa

Observed band size: 45 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMX1b antibody [EPR24053-161] (AB259926)

Immunohistochemical analysis of paraffin-embedded Rat hypothalamus tissue labelling LMX1b with ab259926 at 1/2000 (0.278 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat hypothalamus. The section was incubated with ab259926 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-LMX1b antibody [EPR24053-161] (AB259926)

Immunohistochemical analysis of paraffin-embedded Mouse hypothalamus tissue labelling LMX1b with ab259926 at 1/2000 (0.278 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse hypothalamus (PMID : 27407143). The section was incubated with ab259926 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-LMX1b antibody [EPR24053-161] (AB259926)

Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labelling LMX1b with ab259926 at 1/2000 (0.278 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : almost no staining on mouse stomach. The section was incubated with ab259926 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-LMX1b antibody [EPR24053-161] (AB259926)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labelling LMX1b with ab259926 at 1/2000 (0.278 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on the glomerulus of rat kidney (PMID : 24785190). The section was incubated with ab259926 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-LMX1b antibody [EPR24053-161] (AB259926)

Immunohistochemical analysis of paraffin-embedded Rat E14.5 embryo tissue labelling LMX1b with ab259926 at 1/2000 (0.278 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on the spinal cord of rat E14.5 embryo. The section was incubated with ab259926 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-LMX1b antibody [EPR24053-161] (AB259926)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling LMX1b with ab259926 at 1/2000 (0.278 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on the glomerulus of mouse kidney. The section was incubated with ab259926 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

• WB

Lab

Western blot - Anti-LMX1b antibody [EPR24053-161] (AB259926)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : stomach and ovary (PMID : 24056967).

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 3 minutes

All lanes:

Western blot - Anti-LMX1b antibody [EPR24053-161] (ab259926) at 1/1000 dilution

Lane 1:

Mouse hypothalamus tissue lysate at 40 µg

Lane 2:

Mouse stomach tissue lysate at 40 µg

Lane 3:

Mouse ovary tissue lysate at 40 µg

Lane 4:

Rat hypothalamus tissue lysate at 40 µg

Lane 5:

Rat E17 embryo tissue lysate at 40 µg

Lane 6:

Rat stomach tissue lysate at 40 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 45 kDa

false

• WB

Lab

Western blot - Anti-LMX1b antibody [EPR24053-161] (AB259926)

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

This antibody has a weak cross-reactivity with LMX1a.

Exposure time : 5.5 seconds

All lanes:

Western blot - Anti-LMX1b antibody [EPR24053-161] (ab259926) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with mouse LMX1b full length expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

E.coli extracts containing his-tagged mouse LMX1a full length recombinant protein at 20 µL

Secondary

All lanes:

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

Predicted band size: 42 kDa

false

• Dot

Supplier Data

Dot Blot - Anti-LMX1b antibody [EPR24053-161] (AB259926)

Dot blot analysis of LMX1b using ab259926 at 1 : 1000 (0.556 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Exposure time : 3 minutes

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