Product nameAnti-SMC1 antibody
See all SMC1 primary antibodies
DescriptionRabbit polyclonal to SMC1
Tested applicationsSuitable for: ICC/IF, WB, IP, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat, Chicken, Xenopus laevis
- HeLa and Jurkat whole cell lysate
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab21583 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 150 kDa (predicted molecular weight: 143 kDa).|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
- Information by UniProt
- CDLS2 antibody
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All lanes : Anti-SMC1 antibody (ab21583) at 1 µg/ml
Lane 1 : 20ug HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :
Jurkat whole cell lysate (ab7899) at 20 µg
Lane 3 : 20ug HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with
Human SMC1 peptide (ab23863) at 1 µg/ml
Lane 4 :
Jurkat whole cell lysate (ab7899) at 20 µg with Human SMC1 peptide (ab23863) at 1 µg/ml
All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) (ab28446) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 143 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
ab21583 detects a band of 150 kDa, corresponding to the size of SMC1. This band is competed away by the addition of the immunizing peptide, showing that it is a specific interaction.
SMC1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to SMC1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab21583.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 150kDa: SMC1; Non specific - 41 and 42kDa: We are unsure as to the identity of this extra band.
ICC/IF image of ab21583 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21583, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Panel A shows localisation of ab21583 to the nuclei, Panel B has the Alexa Fluor® 488 channel removed for comparison.
ab21583 staining SMC1 in assynchonous HeLa cells (green). Cells were paraformaldehyde-fixed (4% - 10min) and counterstained with DAPI (red). Secondary antibody: Goat anti-Rabbit conjugated to Cy3 ®. Please refer to Abreview for further details.
IHC image of SMC1 staining in human skin FFPE section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab21583, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
This product has been referenced in:
- Bellelli R et al. Pole Instability Drives Replication Stress, Abnormal Development, and Tumorigenesis. Mol Cell 70:707-721.e7 (2018). Read more (PubMed: 29754823) »
- Zhang Y et al. Phosphorylation of SMC1A promotes hepatocellular carcinoma cell proliferation and migration. Int J Biol Sci 14:1081-1089 (2018). Read more (PubMed: 29988990) »