Anti-SMC3 antibody - ChIP Grade (ab9263)

Immunocytochemistry - SMC3 antibody (ab9263)

SMC3 signals (green) in a human spermatocyte at the zygotene-pachytene transition stage. Wider signals correspond to synapsed areas, whereas narrower signals show non-synapsed regions.

This image was kindly supplied as part of the review submitted by Maria Oliver-Bonet.

Immunocytochemistry - SMC3 antibody (ab9263)

Interphase HeLa cells stained with ab9263 (1/500). The antibody gave predominantly nuclear staining in all interphase cells investigated. ab9263 staining is shown in green. The cells were counterstained with DAPI (red).

Kirk McManus, University of British Columbia

Western blot - SMC3 antibody (ab9263)

All lanes : Anti-SMC3 antibody - ChIP Grade (ab9263) at 1 µg/ml

Lane 1 : Placenta (Human) Tissue Lysate - adult normal tissue (ab29745)
Lane 2 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size : 141.5 kDa
Observed band size : 145 kDa (why is the actual band size different from the predicted?)

Immunoprecipitation - SMC3 antibody (ab9263)

ab9263 Immunoprecipitate in human HeLa cytosolic and nuclear extracts. 500µg of cell lysate incubated with primary antibody (1/100) and matrix (Protein A) in 20mM HEPES ph 7.8, 100mM KAc for 4 hours at 4°C. For Western blotting ab9263 was used at a dilution at 1/1000.

This image is a courtesy of Anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-SMC3 antibody(ab9263)

ab9263 (4µg/ml) staining SMC3 in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining.
Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.