Anti-KIF13B antibody [EPR29559-519]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-KIF13B antibody [EPR29559-519] (AB327425)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized NCI-H929 (human Bone B lymphocyte) and PANC-1(human pancreatic epithelioid carcinoma epithelial cell) cells labelling KIF13B with ab327425 at 1/500 (1.006 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in NCI-H929 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Low expression : PANC-1.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

• I-ELISA

Lab

Indirect ELISA - Anti-KIF13B antibody [EPR29559-519] (AB327425)

Indirect ELISA analysis of ab327425 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Human KIF13A recombinant protein; Human KIF13B recombinant protein.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with Human KIF13A recombinant protein by ELISA.

• WB

Lab

Western blot - Anti-KIF13B antibody [EPR29559-519] (AB327425)

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

Low expression : PANC-1, G-401, skeletal muscle.

The identity of the lower MW band at approximately 130 kDa (in lane 4) is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-KIF13B antibody [EPR29559-519] (ab327425) at 1/1000 dilution

Lane 1:

NCI-H929 (human Bone B lymphocyte) whole cell lysate at 60 µg

Lane 2:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 60 µg

Lane 3:

G-401 (human rhabdoid tumor kidney epithelial cell) whole cell lysate at 60 µg

Lane 4:

Mouse liver tissue lysate at 60 µg

Lane 5:

Mouse skeletal muscle tissue lysate at 60 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 260 kDa,124 kDa

false

Exposure time: 158s

• WB

Lab

Western blot - Anti-KIF13B antibody [EPR29559-519] (AB327425)

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

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-KIF13B antibody [EPR29559-519] (ab327425) at 1/1000 dilution

Lane 1:

A-204 (human muscle rhabdomyosarcoma cell) transfected with scrambled siRNA control whole cell lysate at 80 µg

Lane 2:

A-204 transfected with siRNA specifically targeting KIF13B whole cell lysate at 80 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Observed band size: 260 kDa,124 kDa

false

Exposure time: 6s