Anti-Bim antibody [Y36] - Mouse IgG2a (Chimeric)

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Bim antibody [Y36] - Mouse IgG2a (Chimeric) (AB324516)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labelling Bim with ab324516 at 1/100 (9.4 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in Raji and no staining in WI-38 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : WI-38.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab202272 Recombinant Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab202272) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150117 Goat Anti-Mouse 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-Bim antibody [Y36] - Mouse IgG2a (Chimeric) (AB324516)

Immunohistochemical analysis of paraffin-embedded Panel A Wild-type HAP1 (Human chronic myelogenous leukemia near-haploid cell) cell pellet and Panel B BCL2L11 knockout HAP1 cell pellet labeling Bim with ab324516 at 1/500 (1.874 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (Panel A) wild-type HAP1 cell pellet, no staining on (Panel B) BCL2L11 knockout HAP1 cell pellet. The primary antibody was incubated for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins at room temperature during the LeicaDS9800 kit staining procedure.
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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bim antibody [Y36] - Mouse IgG2a (Chimeric) (AB324516)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Bim with ab324516 at 1/500 (1.874 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human breast cancer. The primary antibody was incubated for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins at room temperature during the LeicaDS9800 kit staining procedure.
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-Bim antibody [Y36] - Mouse IgG2a (Chimeric) (AB324516)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling Bim with ab324516 at 1/500 (1.874 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung cancer. The primary antibody was incubated for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins at room temperature during the LeicaDS9800 kit staining procedure.
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-Bim antibody [Y36] - Mouse IgG2a (Chimeric) (AB324516)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A20 (mouse reticulum sarcoma B lymphocyte) cells labelling Bim with ab324516 at 1/100 (9.4 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in A20 and no staining in B16-F10 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : B16-F10.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab202272 Recombinant Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab202272) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• WB

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Western blot - Anti-Bim antibody [Y36] - Mouse IgG2a (Chimeric) (AB324516)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 24872388).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-4 : 125 seconds; Lane 5 : 37 seconds; Lanes 6-7 : 180 seconds

All lanes:

Western blot - Anti-Bim antibody [Y36] - Mouse IgG2a (Chimeric) (ab324516) at 1/1000 dilution

Lane 1:

Human spleen tissue lysates at 20 µg

Lane 2:

Human thyroid tissue lysates at 20 µg

Lane 3:

Rat spleen tissue lysates at 20 µg

Lane 4:

Rat testis tissue lysates at 20 µg

Lane 5:

Mouse spleen tissue lysates at 20 µg

Lane 6:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 7:

BCL2L11 knockout HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Observed band size: 22 kDa,15 kDa,12 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Bim antibody [Y36] - Mouse IgG2a (Chimeric) (AB324516)

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

Low expression : WI-38 and B16-F10

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Bim antibody [Y36] - Mouse IgG2a (Chimeric) (ab324516) at 1/1000 dilution

Lane 1:

Raji (human Burkitt's lymphoma B lymphocyte) at 20 µg

Lane 2:

WI-38 (human fetal lung fibroblast) at 20 µg

Lane 3:

HEK-293 (human embryonic kidney epithelial cell) at 20 µg

Lane 4:

HeLa (human cervical adenocarcinoma epithelial cell) at 20 µg

Lane 5:

A20 (mouse reticulum sarcoma B lymphocyte) at 20 µg

Lanes 6 - 7:

4T1 (mouse mammary gland carcinoma epithelial cell) at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 22 kDa,15 kDa,12 kDa,36 kDa

false

Exposure time: 180s