Anti-Psoriasin antibody [EPR23482-38] (ab275026) at 1/1000 dilution + Human tonsil tissue lysate at 20 µg

Secondary
VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 11 kDa
Observed band size: 12 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 81 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID: 20596736, 12855640, 26055798).

Immunohistochemical analysis of paraffin-embedded human skin tissue labeling Psoriasin with ab275026 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human skin (PMID: 18223693). The section was incubated with ab275026 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDA-MB-468 cells labelling Psoriasin with ab275026 at 1/250 (2.132 µg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in MDA-MB-468 cells.

Negative control: MDA-MB-231 (PMID: 15994944).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MDA-MB-468 (Human breast adenocarcinoma epithelial cell) cells labelling Psoriasin with ab275026 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Psoriasin was immunoprecipitated from 0.35 mg MDA-MB-468 (human breast adenocarcinoma epithelial cell), whole cell lysate with ab275026 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275026 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: MDA-MB-468 whole cell lysate 10 µg.

Lane 2: ab275026 IP in MDA-MB-468 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275026 in MDA-MB-468 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 76 seconds.

 

All lanes : Anti-Psoriasin antibody [EPR23482-38] (ab275026) at 1/1000 dilution

Lane 1 : His-tagged human recombinant Psoriasin (Protein S100-A7) protein, 500 ng
Lane 2 : His-tagged human recombinant S100-A7A protein, 500 ng

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 11 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 8 seconds.

This antibody reacts with Psoriasin (Protein S100-A7) and Protein S100-A7A.
Linking with 6x His tag leads to the shift of the bands .
Both recombinant proteins were made in house and extracted from E.coli expressing Psoriasin (Protein S100-A7) or Protein S100-A7A.

 

All lanes : Anti-Psoriasin antibody [EPR23482-38] (ab275026) at 1/1000 dilution

Lane 1 : MDA-MB-468 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 11 kDa
Observed band size: 12 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.

The expression profile observed is consistent with what has been described in the literature (PMID: 20596736, 12855640, 26055798).

Negative control: MDA-MB-231 (PMID: 12855640).

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Psoriasin with ab275026 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human tonsil. The section was incubated with ab275026 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.