Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling KIF5A + KIF5B + KIF5C with ab271049 at 1/500 dilution (0.1ug) (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 F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y cells labelling KIF5A + KIF5B + KIF5C with ab271049 at 1/50 (9.72 ug/ml) dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/500 (4 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in SH-SY5Y cell line. is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 (4 ug/ml) dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

KIF5A + KIF5B + KIF5C was immunoprecipitated from 0.35 mg SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 10 ug with ab271049 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271049 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab271049 IP in SH-SY5Y whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271049 in SH-SY5Y whole cell lysate

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

Exposure time : 3 seconds

All lanes:

Immunoprecipitation - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (ab271049)

Observed band size: 117 kDa

false

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C6 (Rat glial tumor glial cell) cells labelling KIF5A + KIF5B + KIF5C with ab271049 at 1/500 dilution (0.1ug) (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, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling KIF5A + KIF5B + KIF5C with ab271049 at 1/50 (9.72 ug/ml) dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/500 (4 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 (4 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labelling KIF5A + KIF5B + KIF5C with ab271049 at 1/500 dilution (0.1ug) (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, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron cells labelling KIF5A + KIF5B + KIF5C with ab271049 at 1/50 (9.72 ug/ml) dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/500 (4 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in rat primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 (4 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 (4 ug/ml) dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

KIF5A + KIF5B + KIF5C was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 10 ug with ab271049 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271049 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 10 ug

Lane 2 : ab271049 IP in Neuro-2a whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271049 in Neuro-2a whole cell lysate

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

Exposure time : 3 seconds

All lanes:

Immunoprecipitation - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (ab271049)

Observed band size: 117 kDa

false

• WB

Lab

Western blot - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

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

Exposure time : 10 seconds

All lanes:

Western blot - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (ab271049) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate at 20 µg

Lane 2:

C2C12 (mouse myoblasts myoblast), whole cell lysate at 20 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

Rat brain cortex tissue lysate at 20 µg

Lane 6:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 117 kDa

Observed band size: 117 kDa

false

• WB

Lab

Western blot - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

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

Degraded fragment (107 KD) has also been described (PMID : 11144355)

Exposure time : 10 seconds

All lanes:

Western blot - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (ab271049) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human hypothalamus tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 117 kDa

Observed band size: 107 kDa,117 kDa

false

• WB

Lab

Western blot - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

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

Degraded fragment (107 KD) has also been described (PMID : 11144355)

Exposure time : 10 seconds

All lanes:

Western blot - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (ab271049) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse cerebral cortex tissue lysate at 20 µg

Lane 3:

Mouse cerebellum tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat cerebellum tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 117 kDa

Observed band size: 107 kDa,117 kDa

false

• WB

Lab

Western blot - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (AB271049)

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

Exposure time : 3 seconds

All lanes:

Western blot - Anti-KIF5A + KIF5B + KIF5C antibody [EPR24325-98] (ab271049) at 1/5000 dilution

Lane 1:

HEK-293T (human embryonic kidney) transfected with KIF5A expression vector containing a myc-His-tag®, whole cell lysate at 2 µg

Lane 2:

HEK-293T transfected with KIF5B expression vector containing a myc-His-tag®, whole cell lysate at 2 µg

Lane 3:

HEK-293T transfected with KIF5C expression vector containing a myc-His-tag®, whole cell lysate at 2 µg

Secondary

All lanes:

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

Predicted band size: 117 kDa

false