Anti-RBPMS antibody [EPR27130-21]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A549 (human lung carcinoma epithelial cell) cells labeling RBPMS with ab309094 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling RBPMS with ab309094 at 1/2000 (0.263 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on rat skeletal muscle. The section was incubated with ab309094 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling RBPMS with ab309094 at 1/2000 (0.263 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cardiac muscle (PMID : 19737887). The section was incubated with ab309094 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling RBPMS with ab309094 at 1/2000 (0.263 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cardiac muscle (PMID : 19737887). The section was incubated with ab309094 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Immunohistochemical analysis of paraffin-embedded Mouse retina tissue labeling RBPMS with ab309094 at 1/2000 (0.263 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on ganglion cell layer of mouse retina (PMID : 24318667). The section was incubated with ab309094 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Immunohistochemical analysis of paraffin-embedded Rat retina tissue labeling RBPMS with ab309094 at 1/2000 (0.263 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on ganglion cell layer of rat retina. The section was incubated with ab309094 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)

Supplier Data

Flow Cytometry (Intracellular) - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 3T3-L1 (mouse embryonic fibroblast) cells labeling RBPMS with ab309094 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse retina tissue staining RBPMS with ab309094 at a 1/2000 dilution, ab140603 anti-CPX used at 1/2000 dilution and ab259971 anti-GAT1 used at a 1/500 dilution.

Panel A : merged staining of anti-RBPMS (green; Opal™520), anti-CPX (magenta; Opal™690) and anti-GAT1 (gray; Opal™570) on mouse retina.

Panel B : anti-RBPMS staining ganglion cell layer in mouse retina.

Panel C : ant-CPX staining outer nuclear layer and inner nuclear layer in mouse retina.

Panel D : ant-GAT1 staining inner plexiform layer in mouse retina.

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

The section was incubated in three rounds of staining : in the order of ab309094, ab140603 and ab259971 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat retina tissue staining RBPMS with ab309094 at a 1/2000 dilution, ab140603 anti-CPX used at 1/2000 dilution and ab259971 anti-GAT1 used at a 1/500 dilution.

Panel A : merged staining of anti-RBPMS (green; Opal™520), anti-CPX (magenta; Opal™690) and anti-GAT1 (gray; Opal™570) on rat retina.

Panel B : anti-RBPMS staining ganglion cell layer in rat retina.

Panel C : ant-CPX staining outer nuclear layer and inner nuclear layer in rat retina.

Panel D : ant-GAT1 staining inner plexiform layer in rat retina.

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

The section was incubated in three rounds of staining : in the order of ab309094, ab140603 and ab259971 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 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-RBPMS antibody [EPR27130-21] (AB309094)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling RBPMS with ab309094 at 1/2000 (0.263 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse skeletal muscle. The section was incubated with ab309094 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.

• WB

Lab

Western blot - Anti-RBPMS antibody [EPR27130-21] (AB309094)

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

All lanes:

Western blot - Anti-RBPMS antibody [EPR27130-21] (ab309094) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 6:

C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

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 1/20000 dilution

Observed band size: 25 kDa,30 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-RBPMS antibody [EPR27130-21] (AB309094)

Blocking and diluting buffer and concentration : 5% NFDM /TBST. No bands were detected by antibody ab309094 in 20μg retinal tissue lysates. For samples with low RBPMS protein expression levels, we recommend : 1. Increasing lysate load to 50μg 2. Using a lower antibody dilution 3. Using femtogram-level sensitivity substrates

All lanes:

Western blot - Anti-RBPMS antibody [EPR27130-21] (ab309094) at 1/1000 dilution

Lane 1:

Human heart tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse retina tissue lysate at 20 µg

Lane 5:

Rat heart tissue lysate at 20 µg

Lane 6:

Rat kidney tissue lysate at 20 µg

Lane 7:

Rat retina tissue lysate at 20 µg

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 1/20000 dilution

Observed band size: 25 kDa,30 kDa

false

Exposure time: 180s