Immunohistochemistry of mouse testis cells labeling XBP1 with Anti-XBP1 antibody (ab37152) at 2μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4^oC. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry of human liver cells labeling XBP1 with Anti-XBP1 antibody (ab37152) at 20μg/ml. followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green). DAPI staining (Blue).

Immunohistochemistry of mouse spleen cells labeling XBP1 with Anti-XBP1 antibody (ab37152) at 2μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4^oC. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry of mouse pancreas cells labeling XBP1 with Anti-XBP1 antibody (ab37152) at 20μg/ml. followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green). DAPI staining (Blue).

Lane 1 : Anti-XBP1 antibody (ab37152) at 1 µg/ml
Lane 2 : Anti-XBP1 antibody (ab37152) at 2 µg/ml

All lanes : HepG2 cell lysate

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

Predicted band size: 29 kDa

Immunohistochemistry of human pancreas cells labeling XBP1 with Anti-XBP1 antibody (ab37152) at 20μg/ml. followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green). DAPI staining (Blue).

Lane 1 : Anti-XBP1 antibody (ab37152) at 1 µg/ml
Lane 2 : Anti-XBP1 antibody (ab37152) at 2 µg/ml
Lane 3 : Anti-XBP1 antibody (ab37152) at 4 µg/ml

All lanes : HepG2 cell lysate

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : goat anti-rabbit HRP at 1/10000 dilution

Predicted band size: 29 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?

Gel: 10%

Running condition: 130V, 1.5hrs

Transfer condition: Wet, 250mA, 2 hrs (Nitrocellulose membrane)

Blocking condition: 5% non-fat milk in TBS at 4 overnight

Primary antibody incubation: Room temperature for 1 hour

Secondary antibody incubation: Room temperature for 1 hour

Washing condition: 5 mL TBST, 4 x 5 minutes

Exposure system: ECL 

Immunocytochemistry/Immunofluorescence of HepG2 cells labeling XBP1 with Anti-XBP1 antibody (ab37152) at 20μg/ml. Blue: DAPI staining.

 

Immunohistochemistry (Formalin-fixed paraffin embedded sections) of human liver tissue labeling XBP1 with Anti-XBP1 antibody (ab37152) at 5μg/ml.

ab37152 staining XBP1 in human liver tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using Citrate-EDTA antigen retrieval buffer and microwave. Heating was performed in the microwave 3 minutes on high until boiling, 6 minutes on 20% power. Samples were then blocked for 15 minutes at 27°C followed by incubation with the primary antibody at a 1/100 dilution for 12 hours at 4°C. An undiluted HRP-conjugated rat polyclonal was used as the secondary antibody.

See Abreview

ab37152 at 1/200 dilution staining XBP1 in human non-lesional skin tissue sections by Immunohistochemistry (Formalin/ PFA fixed paraformaldehyde tissue sections). A biotin labelled secondary was used at 1/100 dilution. Diaminobenzidine (DAB) was used for staining development in a ABC staining Kit and the sections were counterstained with haematoxylin.

Immunocytochemistry of XBP-1 in HepG2 cells with ab37152 at 10 ug/mL