Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeing ASH2L with unpurified ab176334 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

Immunofluorescent analysis of HeLa labeling ASH2L with ab176334 at 1/100 dilution.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

Immunocytochemistry/ Immunofluorescence analysis of 293T (Human embryonic kidney epithelial cell) cells labeling ASH2L with purified ab176334 at 1/50 dilution (7.78 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) 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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ASH2L (red) with unpurifiedab176334 at a 1/1200 dilution. Cells were fixed with 80% methanol and permeabilized with 0.1% Tween-20. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeing ASH2L with unpurified ab176334 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue sections labeling ASH2L with purified ab176334 at 1/8000 dilution (0.05 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling ASH2L with purified ab176334 at 1/8000 dilution (0.05 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling ASH2L with purified ab176334 at 1/8000 dilution (0.05 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Lab

Western blot - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

ab176334 recoginzes 3 isoforms of ASH2L

All lanes:

Western blot - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (ab176334) at 1/10000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

Mouse heart lysates at 20 µg

Lane 4:

Rat heart lysates 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: 69 kDa

Observed band size: 65 kDa,80 kDa

false

• WB

Supplier Data

Western blot - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

All lanes:

Western blot - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (ab176334) at 1/10000 dilution

Lane 1:

HeLa cell lysates at 10 µg

Lane 2:

293T cell lysates at 10 µg

Lane 3:

Jurkat cell lysates at 10 µg

Lane 4:

K562 cell lysates at 10 µg

Predicted band size: 69 kDa

false

• WB

CiteAb

Western blot - Anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker (AB176334)

ASH2L western blot using anti-ASH2L antibody [EPR13107(B)] - Nuclear Marker ab176334. Publication image and figure legend from Rao, R. A., Ketkar, A. A., et al., 2019, EMBO Rep, PubMed 30858340.

ab176334 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab176334 please see the product overview.

EHMT1 interacts with LMNB1 domain and co‐regulate a subset of genesAEHMT1 interacts with EHMT2, LMNB1, and HP1. Whole cell lysates from fetal HDFs were subjected to IP reaction using EHMT1 or LMNB1 antibody. Subsequently, Western blot was performed using the IPed material to detect EHMT1, EHMT2, LMNB1, and HP1. ASH2L did not show any interaction with EHMT1 and LMNB1 thus acted as a negative control.BEHMT1 interacts with LMNB1 and EHMT2 via SET domain. HEK293 cells were transfected with pEGFP‐Ankyrin (pEGFPC1‐ANK) or pEGFPC1‐SET domains of EHMT1 to determine domain‐specific association with LMNB1. Cell extracts were subjected to IP using GFP‐antibody, and the bound complexes were then analyzed by immunoblot using LMNB1, EHMT2, and GFP antibodies. HEK‐293 whole cell extract represents 1% input. pEGFPC1 empty vector transfected HEK293 or untransfected HEK293 cells were used as control reactions. Arrows indicate specific band.CCoomassie‐stained SDS–PAGE gel showing 6X His EHMT1‐SET purified protein used for methyltransferase assays.D–FGene ontology (GO) analysis of EHMT1 and LMNB1 bound genes. Representative figure showing enriched GO terms for EHMT1 (D), LMNB1 (E), and EHMT1 and LMNB1 (F) co‐bound genes. The length of the bar (y‐axis) denotes total genes falling within GO term.GCircos plot showing genome wide peak density of EHMT1 (green) and LMNB1 (red).

false