All lanes : Anti-GBP1 antibody [1B1] (ab252832) at 1/1000 dilution

Lane 1 : Untreated HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : HeLa treated with 100U/ml IFN gamma (ab9659) for 24 hours, whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 68 kDa
Observed band size: 67 kDa why is the actual band size different from the predicted?


Exposure time: 3 seconds

The expression profile observed is consistent with what has been described in the literature (PMID:19079332)

Blocking/diluting buffer and concentration: 5% NFDM/TBST

Formalin-fixed, paraffin-embedded human spleen tissue stained for GBP1 using ab252832 at 1/100 dilution in immunohistochemical analysis followed by a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Cytoplasmic staining on human spleen. The section was incubated with ab252832 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. 

Secondary antibody only control: Used PBS instead of primary antibody. Secondary antibody was a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

Formalin-fixed, paraffin-embedded human tonsil tissue stained for GBP1 using ab252832 at 1/100 dilution in immunohistochemical analysis followed by a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Cytoplasmic staining on human tonsil. The section was incubated with ab252832 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. 

Secondary antibody only control: Used PBS instead of primary antibody. Secondary antibody was a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

ICC/IF analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (+/-IFN gamma (100U/ml 24h)) cells labeling GBP1 (Green) using ab252832 at 1/30 dilution followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution. Tubulin was counterstained using ab179504  Anti-beta IV Tubulin at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/1000 diluton (Red). The nuclear counterstain was DAPI (Blue). Confocal image showing cytoplasmic staining in HeLa cells treated with interferon gamma (100 U/ml) for 24 hours.