Anti-KPNB1 antibody

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KPNB1 antibody (AB45938)

IHC image of KPNB1 staining in a formalin fixed, paraffin embedded normal human testis tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab45938, 5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IP

Unknown

Immunoprecipitation - Anti-KPNB1 antibody (AB45938)

KPNB1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5μg of Rabbit polyclonal to KPNB1 and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70^oC; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab45938.
Secondary : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band : 97kDa; KPNB1

All lanes:

Immunoprecipitation - Anti-KPNB1 antibody (ab45938)

Predicted band size: 97 kDa

false

• WB

Project

Western blot - Anti-KPNB1 antibody (AB45938)

All lanes:

Western blot - Anti-KPNB1 antibody (ab45938) at 2 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 15 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 15 µg

Lane 3:

Western blot - NIH/3T3 whole cell lysate (<a href='/en-us/products/cell-lysates/nih-3t3-whole-cell-lysate-ab7179'>ab7179</a>) at 15 µg

Lane 4:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 15 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 97 kDa

Observed band size: 97 kDa

false