Western blot - SOCS3 antibody (ab16030)

All lanes : Anti-SOCS3 antibody (ab16030) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate
Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with SOCS3 peptide (ab16199) at 1 µg/ml
Lane 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with SOCS3 peptide (ab16199) at 1 µg/ml
Lane 7 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with SOCS3 peptide (ab16199) at 1 µg/ml
Lane 8 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate (ab52252) with SOCS3 peptide (ab16199) at 1 µg/ml

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.

Observed band size : 30 kDa (why is the actual band size different from the predicted?)


The 30kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to SOCS3.

Immunocytochemistry/ Immunofluorescence - SOCS3 antibody (ab16030)

ICC/IF image of ab16030 stained human HeLa cells. The cells were PFA fixed (10 min) and incubated with the antibody (ab16030, 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-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (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).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - SOCS3 antibody (ab16030)

Image courtesy of Human Protein Atlas

ab16030 staining SOCS3 in human bone marrow showing strong staining within the bone marrow. Paraffin embedded human bone marrow was incubated with ab16030 (1/100) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH6.

ab16030 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

Immunohistochemistry (Frozen sections) - SOCS3 antibody (ab16030)

ab16030 staining SOCS3 in Human pancreatic tumour tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/300 in PBS) for 8 hours at 4ºC. An Alexa Fluor^®555-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody. Nuclei were stained by DAPI.

This image is courtesy of an anonymous Abreview