IHC-P - PLU1 / Jarid1B antibody (ab56759)

PLU1 / Jarid1B antibody (ab56759) used in immunohistochemistry at 1ug/ml on formalin fixed and paraffin embedded human colon.

Western blot - KDM5B / PLU1 / Jarid1B antibody (ab56759)

Predicted band size : 190 kDa


PLU1 / Jarid1B antibody (ab56759) at 1ug/lane + HeLa cell lysate at 25ug/lane.

Western blot - KDM5B / PLU1 / Jarid1B antibody (ab56759)

All lanes : Anti-KDM5B / PLU1 / Jarid1B antibody (ab56759) at 1/500 dilution

Lane 1 : Human Pancreatic Cancer whole cell lysate (Line AsPc1)
Lane 2 : Human Pancreatic Cancer whole cell lysate (line Panc1)
Lane 3 : Human Pancreatic Cancer whole cell lysate (Line CaPan1)

Lysates/proteins at 35 µg per lane.

Secondary
An HRP-conjugated Sheep anti-mouse polyclonal at 1/5000 dilution
developed using the ECL technique

Performed under non-reducing conditions.

Predicted band size : 190 kDa
Observed band size : 190 kDa


Exposure time : 15 minutes

Blocking Step: 10% Milk for 1 hour at room temperature

This image is courtesy of an Abreview submitted by Pawel Mazur

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - KDM5B / PLU1 / Jarid1B antibody (ab56759)

ab56759 staining KDM5B/PLU1/Jarid1B in Human pancreatic cancer tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with PBS + Triton 0.025% and blocked with 10% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4ºC. A Biotin-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence-KDM5B / PLU1 / Jarid1B antibody(ab56759)

ICC/IF image of ab56759 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 (ab26759, 1µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

Flow Cytometry-Anti-KDM5B / PLU1 / Jarid1B antibody(ab56759)

Overlay histogram showing MCF7 cells stained with ab56759 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56759, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.