Anti-KLB antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLB antibody (AB106794)

IHC image of KLB staining in Human normal stomach formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab106794, 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.

• WB

Unknown

Western blot - Anti-KLB antibody (AB106794)

KLB contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-KLB antibody (ab106794) at 1 µg/mL

All lanes:

HepG2 whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 119 kDa

Observed band size: 125 kDa,39 kDa,54 kDa

true

Exposure time: 2min

• WB

Lab

Western blot - Anti-KLB antibody (AB106794)

Western blot : Rabbit Polyclonal to KLB ab106794 staining at 1 µg/mL, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 120 kDa in Wild-type A549 cell lysates with no signal observed at this size in KLB knockout A549 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-KLB antibody (ab106794) at 1 µg/mL

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

KLB knockout A549 at 20 µg

Lane 3:

HepG2 at 20 µg

Lane 4:

THP-1 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 119.808 kDa

Observed band size: 120 kDa

false