Anti-Menin antibody [EPR3986]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Menin antibody [EPR3986] (AB92443)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue sections labelling Menin with purified ab92443 at 1/500 dilution (1.22 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Menin antibody [EPR3986] (AB92443)

Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Menin with purified ab92443 at 1/50 dilution (12.1 μg/mL). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.6 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150078) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IP

Lab

Immunoprecipitation - Anti-Menin antibody [EPR3986] (AB92443)

Purified ab92443 at 1/50 dilution (2μg) immunoprecipitating Menin in Jurkat whole cell lysate.
Lane 1 (input) : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10μg
Lane 2 (+) : ab92443 + Jurkat whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab92443 in Jurkat whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 75 kDa

All lanes:

Immunoprecipitation - Anti-Menin antibody [EPR3986] (ab92443)

Predicted band size: 68 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Menin antibody [EPR3986] (AB92443)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labelling Menin with purified ab92443 at 1/100 dilution (6.08 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Menin antibody [EPR3986] (AB92443)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labelling Menin with purified ab92443 at 1/100 dilution (6.08 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

• IP

Supplier Data

Immunoprecipitation - Anti-Menin antibody [EPR3986] (AB92443)

Menin was immunoprecipitated from 0.35 mg MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate 10 ug with ab92443 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab92443 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate 10 μg

Lane 2 : ab92443 IP in MEF whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab92443 in MEF whole cell lysate

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

Exposure time : 3 minutes

All lanes:

Immunoprecipitation - Anti-Menin antibody [EPR3986] (ab92443)

Predicted band size: 68 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-Menin antibody [EPR3986] (AB92443)

All lanes:

Western blot - Anti-Menin antibody [EPR3986] (ab92443) at 1/1000 dilution

Lane 1:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 15 µg

Lane 2:

MEF (Mouse embryonic fibroblast (immortalized)) whole cell lysate at 15 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate at 15 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: 68 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-Menin antibody [EPR3986] (AB92443)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : Menin knockout HAP1 cell lysate (20 μg)
Lane 3 : Jurkat cell lysate (20 μg)
Lane 4 : A431 cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab92443 observed at 74 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab92443 was shown to recognize Menin when Menin knockout samples were used, along with additional cross-reactive bands. Wild-type and Menin knockout samples were subjected to SDS-PAGE. ab92443 and ab8245 (loading control to GAPDH) were diluted 1/10,000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Menin antibody [EPR3986] (ab92443)

Predicted band size: 68 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Menin antibody [EPR3986] (AB92443)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 K-562 (human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab92443 [EPR3986]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Menin antibody [EPR3986] (AB92443)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 K-562 (human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab92443 [EPR3986]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Menin antibody [EPR3986] (AB92443)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 K-562 (human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab92443 [EPR3986]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.