Anti-Mre11 antibody [EPR21027] - ChIP Grade (ab208020) at 1/1000 dilution + Human fetal kidney tissue lysate at 10 µg

Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

Developed using the ECL technique.

Predicted band size: 81 kDa
Observed band size: 81 kDa

Blocking/Dilution: 5% NFDM/TBST.

Exposure time: 3 minutes.

All lanes : Anti-Mre11 antibody [EPR21027] - ChIP Grade (ab208020) at 1/5000 dilution

Lane 1 : K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lane 2 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 4 : D3 (mouse embryo stem cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 81 kDa
Observed band size: 81 kDa

Blocking/Dilution: 5% NFDM/TBST.

Exposure time: 3 minutes.

All lanes : Anti-Mre11 antibody [EPR21027] - ChIP Grade (ab208020) at 1/1000 dilution

Lane 1 : C6 (rat glial tumor cell line) whole cell lysate
Lane 2 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 5 : Mouse brain tissue lysate
Lane 6 : Rat brain tissue lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 81 kDa
Observed band size: 81 kDa

Blocking/Dilution: 5% NFDM/TBST.

Exposure time: 3 minutes.

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Mre11 with ab208020 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use. Nuclear staining in human colon tissue is observed (PMID: 25447316). Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue labeling Mre11 with ab208020 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use. Nuclear staining in human endometrial carcinoma is observed (PMID: 24927325). Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

 

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Mre11 with ab208020 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use. Nuclear staining in mouse testis is observed (PMID: 10508516). Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

 

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Mre11 with ab208020 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use. Nuclear staining in rat colon tissue is observed (PMID: 28357369). Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

 

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized K562 (Human human chronic myelogenous leukemia cell line from bone marrow) cell line labeling Mre11 with ab208020 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

Chromatin was prepared from non-treated HeLa cells or HeLa cells treated with 1% Methyl methanesulfonate for 1 hour according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 5 µg of ab208020 (blue), and 20 µL of protein A/G sepharose beads slurry (10 µL of sepharose A beads + 10 µL of sepharose G beads). 5 μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).