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Synthetic peptide (Human) conjugated to KLH - which represents a portion within the last 100 amino acids of the human Structural Maintenance of Chromosomes-1 (GenBank PID 5453642).
Our Abpromise guarantee covers the use of ab9262 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC||1/200 - 1/800.|
|IP||Use at 1-4 µg/mg of lysate.|
|WB||1/1000 - 1/10000. Detects a band of approximately 160 kDa (predicted molecular weight: 143 kDa).|
|ChIP||Use at an assay dependent concentration. PubMed: 20966046|
Interphase HeLa cells stained with ab9262 (1/500). The antibody gave nuclear staining in all interphase nuclei investigated. However, the antibody failed to recognize any chromatin-associated epitopes in prophase or metaphase cells suggesting that the epitopes may be masked during mitosis. ab9262 staining is shown in green. The cells were counterstained with DAPI (red). 100x magnification.
Immunoprecipitation analysis of HeLa whole cell lysate (0.5 or 1.0 mg per IP reaction; 20% of IP loaded) prepared using NETN lysis buffer.
Lanes 1 & 2: ab9262 was used for IP at 6 µg per reaction.
Lane 3: IP using rabbit anti-SMC1A antibody (ab140493).
Lane 4: Control IgG.
For western blotting immunoprecipitated SMC1, ab9262 was used at 1 µg/ml.
Ab9262 staining human normal colon tissue. Staining is localised to nuclear compartment.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
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