Anti-CARM1 antibody [EPR23678-113]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CARM1 antibody [EPR23678-113] (AB243638)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CARM1 with ab243638 at 1/200 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon (PMID : 20462455). The section was incubated with ab243638 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 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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CARM1 antibody [EPR23678-113] (AB243638)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling CARM1 with ab243638 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CARM1 antibody [EPR23678-113] (AB243638)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling CARM1 with ab243638 at 1/200 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon carcinoma (PMID : 20462455). The section was incubated with ab243638 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 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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CARM1 antibody [EPR23678-113] (AB243638)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Wild-type HEK-293T (human embryonic kidney epithelial cell, Right panel)/ CARM1 knockout HEK-293T cells (Left panel) cells labeling CARM1 with ab243638 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Positive staining on HEK-293T cells (ab255449), while no staining on CARM1 knockout HEK-293T cells (ab266557).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CARM1 antibody [EPR23678-113] (AB243638)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling CARM1 with ab243638 at 1/200 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse liver. The section was incubated with ab243638 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 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-CARM1 antibody [EPR23678-113] (AB243638)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling CARM1 with ab243638 at 1/200 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat colon. The section was incubated with ab243638 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 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-CARM1 antibody [EPR23678-113] (AB243638)

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

Samples are non-boiled as boiling may cause protein aggregates

Exposure time : 10 seconds

All lanes:

Western blot - Anti-CARM1 antibody [EPR23678-113] (ab243638) at 1/1000 dilution

Lane 1:

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

Lane 2:

HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

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 1/100000 dilution

Predicted band size: 65 kDa

Observed band size: 65 kDa

false

• WB

Lab

Western blot - Anti-CARM1 antibody [EPR23678-113] (AB243638)

Blocking and diluting buffer and concentration : Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

Lanes 1 - 4 : Merged signal (red and green). Green - ab243638 observed at 65 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

ab243638 was shown to react with CARM1 in wild-type HEK-293T cells in western blot with loss of signal observed in CARM1 knockout cell line ab266557 (CARM1 knockout cell lysate ab257871). Wild-type and CARM1 knockout HEK-293T cell lysates were subjected to SDS-PAGE.

ab243638 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

Boiled samples are used in this blot.

All lanes:

Western blot - Anti-CARM1 antibody [EPR23678-113] (ab243638) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 2:

CARM1 knockout HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 65 kDa

Observed band size: 65 kDa

false

• WB

Lab

Western blot - Anti-CARM1 antibody [EPR23678-113] (AB243638)

Blocking and diluting buffer and concentration : Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

Lanes 1 - 4 : Merged signal (red and green). Green - ab243638 observed at 65 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

ab243638 was shown to react with CARM1 in wild-type HEK-293T cells in western blot with loss of signal observed in CARM1 knockout cell line ab266557 (CARM1 knockout cell lysate ab257871). Wild-type and CARM1 knockout HEK-293T cell lysates were subjected to SDS-PAGE.

ab243638 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

Boiled samples are used in this blot.

All lanes:

Western blot - Anti-CARM1 antibody [EPR23678-113] (ab243638) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 2:

CARM1 knockout HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 2:

Western blot - Human CARM1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-carm1-knockout-hek-293t-cell-line-ab266557'>ab266557</a>)

Lane 3:

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

Lane 4:

HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 65 kDa

Observed band size: 65 kDa

false

• WB

Lab

Western blot - Anti-CARM1 antibody [EPR23678-113] (AB243638)

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

Samples are non-boiled as boiling may cause protein aggregates

Exposure times :

Lane 1-2 : 15 seconds

Lane 3 : 92 seconds

All lanes:

Western blot - Anti-CARM1 antibody [EPR23678-113] (ab243638) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Rat liver tissue 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 1/100000 dilution

Predicted band size: 65 kDa

Observed band size: 65 kDa

false