Anti-ROR alpha/RORA antibody [EPR23719-18] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-ROR alpha/RORA antibody [EPR23719-18] - BSA and Azide free (AB278108)

This data was developed using ab256799, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat cerebellum tissue staining SOX1 with ab242125 at a 1/2000 dilution.

Panel A : merged staining of anti-SOX1 (green; Opal^™570), anti-SOX10 (magenta; Opal^™690) and anti-ROR alpha (gray; Opal^™520) on rat cerebellum.

Panel B : anti-SOX1 staining Bergmann glia in rat cerebellum.

Panel C : anti-SOX10 staining glial cells in rat cerebellum.

Panel D : anti-ROR alpha staining Purkinje cell and molecular layers in rat cerebellum.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab242125, ab180862 and ab256799 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system using Opal Polymer HRP Ms + Rb (ready to use).

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with a Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed 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-ROR alpha/RORA antibody [EPR23719-18] - BSA and Azide free (AB278108)

This data was developed using ab256799, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling ROR alpha/RORA with ab256799 at 1/800 (0.601 μg/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse cerebellum ( PMID : 14966198). The section was incubated with ab256799 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 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-ROR alpha/RORA antibody [EPR23719-18] - BSA and Azide free (AB278108)

This data was developed using ab256799, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling ROR alpha/RORA with ab256799 at 1/800 (0.601 μg/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat cerebellum. The section was incubated with ab256799 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-ROR alpha/RORA antibody [EPR23719-18] - BSA and Azide free (AB278108)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256799).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebellum tissue labelling SOX1 with ab242125 at 1 : 2000 (0.29 ug/ml) dilution, RoR alpha/RORA with ab256799 at 1 : 800 (0.63 ug/ml) dilution and SOX10 with ab200078 at 1 : 5000 (0.2 ug/ml) followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-SOX1 (magenta; Opal^™690), anti-RORA (green; Opal^™520) and anti-SOX10 (gray; Opal^™570) on mouse cerebellum.
Panel B : anti-SOX1 staining Bergmann glia in mouse cerebellum.
Panel C : anti-RORA staining Pukinje cell and molecular layers in mouse cerebellum.
Panel D : anti-SOX10 staining glial cells in mouse cerebellum.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab242125, ab256799 and ab200078 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] - BSA and Azide free (AB278108)

This data was developed using ab256799, the same antibody clone in a different buffer formulation.

ROR alpha/RORA was immunoprecipitated from 0.35 mg rat cerebellum tissue lysate with ab256799 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256799 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Rat cerebellum tissue lysate 10μg

Lane 2 : ab256799 IP in rat cerebellum tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab256799 in rat cerebellum tissue lysate

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

Exposure time : 3 minutes

All lanes:

Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] (<a href='/ko/products/primary-antibodies/ror-alpha-rora-antibody-epr23719-18-ab256799'>ab256799</a>)

Predicted band size: 63 kDa

Observed band size: 60 kDa,65 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] - BSA and Azide free (AB278108)

This data was developed using ab256799, the same antibody clone in a different buffer formulation.

ROR alpha/RORA was immunoprecipitated from 0.35 mg mouse cerebellum tissue lysate with ab256799 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256799 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse cerebellum tissue lysate 10μg

Lane 2 : ab256799 IP in mouse cerebellum tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab256799 in mouse cerebellum tissue lysate

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

Exposure time : 3 minutes

All lanes:

Immunoprecipitation - Anti-ROR alpha/RORA antibody [EPR23719-18] (<a href='/ko/products/primary-antibodies/ror-alpha-rora-antibody-epr23719-18-ab256799'>ab256799</a>)

Predicted band size: 63 kDa

Observed band size: 60 kDa,65 kDa

false

• WB

Lab

Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] - BSA and Azide free (AB278108)

This data was developed using ab256799, the same antibody clone in a different buffer formulation.

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

Low expression : mouse spleen (PMID : 9226375).

The molecular weight observed is consistent with what has been described in the literature (PMID : 19955433, PMID : 12023888).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] (<a href='/ko/products/primary-antibodies/ror-alpha-rora-antibody-epr23719-18-ab256799'>ab256799</a>) at 1/1000 dilution

Lane 1:

Mouse cerebellum tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 63 kDa

Observed band size: 60 kDa,65 kDa

false

• WB

Lab

Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] - BSA and Azide free (AB278108)

This data was developed using ab256799, the same antibody clone in a different buffer formulation.

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

Exposure time : 3 minutes

All lanes:

Western blot - Anti-ROR alpha/RORA antibody [EPR23719-18] (<a href='/ko/products/primary-antibodies/ror-alpha-rora-antibody-epr23719-18-ab256799'>ab256799</a>) at 1/1000 dilution

All lanes:

Rat cerebellum tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 63 kDa

Observed band size: 60 kDa,65 kDa

false