Anti-SETD1B antibody [EPR25142-11]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SETD1B antibody [EPR25142-11] (AB300479)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling SETD1B with ab300479 at 1/50 dilution (1ug) (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 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SETD1B antibody [EPR25142-11] (AB300479)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labelling SETD1B with ab300479 at 1/50 dilution (1ug) (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 1/2000 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-SETD1B antibody [EPR25142-11] (AB300479)

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

Exposure time : Lane1 : 3 minutes, Lane 2 : 26 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29138278).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-SETD1B antibody [EPR25142-11] (ab300479) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

F9 (mouse embryonal carcinoma 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

Predicted band size: 208 kDa

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• WB

Lab

Western blot - Anti-SETD1B antibody [EPR25142-11] (AB300479)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29138278).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-SETD1B antibody [EPR25142-11] (ab300479) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa transfected with siRNA specifically targeting SETD1B, 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

Predicted band size: 209 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-SETD1B antibody [EPR25142-11] (AB300479)

Exposure time : Lane 1 - 3 minutes, Lane 2 : 26 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29138278).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

An unknown band at ~115 kDa was detected in lane 3.

All lanes:

Western blot - Anti-SETD1B antibody [EPR25142-11] (ab300479) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma) 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

Predicted band size: 209 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SETD1B antibody [EPR25142-11] (AB300479)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling SETD1B with ab300479 at 1/100 (5.2 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in HeLa cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/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 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SETD1B antibody [EPR25142-11] (AB300479)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labelling SETD1B with ab300479 at 1/100 (5.2 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in F9 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/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 2 ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SETD1B antibody [EPR25142-11] (AB300479)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling SETD1B with ab300479 at 1/100 (5.2 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Nuclear staining on human colon at a low level (PMID : 24925220). The section was incubated with ab300479 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SETD1B antibody [EPR25142-11] (AB300479)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling SETD1B with ab300479 at 1/100 (5.2 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Nuclear staining on human colon cancer (PMID : 24925220). The section was incubated with ab300479 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SETD1B antibody [EPR25142-11] (AB300479)

Immunohistochemical analysis of paraffin-embedded Mouse liver cancer tissue labeling SETD1B with ab300479 at 1/100 (5.2 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Nuclear staining on mouse liver cancer. The section was incubated with ab300479 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)