Anti-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling RG9MTD1 with ab313372 at 1/500 dilution (0.1 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-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling RG9MTD1 with ab313372 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Punctate staining on human colon.The section was incubated with ab313372 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-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling RG9MTD1 with ab313372 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Punctate staining on human kidney.The section was incubated with ab313372 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-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon carcinom tissue labeling RG9MTD1 with ab313372 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Punctate staining on human colon carcinoma.The section was incubated with ab313372 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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling RG9MTD1 with ab313372 at 1/50 (10.28 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing mitochondrial staining in A431 cells.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab33985 Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/1000 1ug/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 2ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling RG9MTD1 with ab313372 at 1/50 (10.28 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing mitochondrial staining in HeLa cells.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8) . ab33985 Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/1000 1ug/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 2ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling RG9MTD1 with ab313372 at 1/500 dilution (0.1 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.

• IP

Supplier Data

Immunoprecipitation - Anti-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. RG9MTD1 was immunoprecipitated from 0.35 mg A431 (human epidermoid carcinoma epithelial cell) whole cell lysate with ab313372 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313372 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : A431 (human epidermoid carcinoma epithelial cell) whole cell lysate Lane 2 : ab313372 IP in A431 (human epidermoid carcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313372 in A431 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 seconds

All lanes:

Immunoprecipitation - Anti-RG9MTD1 antibody [EPR27065-61] (<a href='/en-us/products/primary-antibodies/rg9mtd1-antibody-epr27065-61-ab313372'>ab313372</a>) at 1/30 dilution

All lanes:

A431 (human epidermoid carcinoma epithelial cell) whole cell 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: 3s

• WB

Supplier Data

Western blot - Anti-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200, 000 dilution. Exposure time : 3 seconds

All lanes:

Western blot - Anti-RG9MTD1 antibody [EPR27065-61] (<a href='/en-us/products/primary-antibodies/rg9mtd1-antibody-epr27065-61-ab313372'>ab313372</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeti RG9MTD1 whole cell lysate at 20 µg

Lane 3:

A431 (human epidermoid 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:

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

Lane 6:

Huh7 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell 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 1/20000 dilution

Observed band size: 44 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-RG9MTD1 antibody [EPR27065-61] - BSA and Azide free (AB313373)

This data was developed using ab313372, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lanes 1-2 : 15 seconds; Lanes 3-4 : 26 seconds.

All lanes:

Western blot - Anti-RG9MTD1 antibody [EPR27065-61] (<a href='/en-us/products/primary-antibodies/rg9mtd1-antibody-epr27065-61-ab313372'>ab313372</a>) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell 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 1/20000 dilution

Observed band size: 44 kDa

false