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ab75025
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Reassurance, Refunds & Replacements
If your product does not perform as described on this datasheet, we will refund or replace your product...
Read our guarantee »This product is covered by the Abpromise guarantee. Our scientific support team are available to answer any questions or queries - fill out an inquiry form for ab49573 for help.
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Western blot - SIGIRR antibody (ab49573)
All lanes : Anti-SIGIRR antibody (ab49573) at 1 µg/ml
Lane 1 : Kidney (Mouse) Tissue Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : SK N SH (Human neuroblastoma) 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
Performed under reducing conditions.
Predicted band size : 46 kDa
Observed band size : 48 kDa (why is the actual band size different from the predicted?)
Additional bands at : 27 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 6 minutes
SIGIRR contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. The 27 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to SIGIRR.
Immunocytochemistry/ Immunofluorescence - SIGIRR antibody (ab49573)
ICC/IF image of ab49573 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab49573, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, hepG2 and MCF7 cells at 5µg/ml.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - SIGIRR antibody (ab49573)
IHC image of SIGIRR staining in Human Normal Liver FFPE section, performed on a 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 ab49573, 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.
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