Anti-KIF5B+KIF5C antibody [EPR10276(B)] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-KIF5B+KIF5C antibody [EPR10276(B)] - BSA and Azide free (AB232350)

Immunofluorescence analysis of HeLa cells, labeling KIF5B using unpurified ab167429 at a 1/100 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167429).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIF5B+KIF5C antibody [EPR10276(B)] - BSA and Azide free (AB232350)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human brain tissue, labeling KIF5B using unpurified ab167429 at a 1/100 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167429).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-KIF5B+KIF5C antibody [EPR10276(B)] - BSA and Azide free (AB232350)

Immunocytochemistry/Immunofluorescence analysis HepG2(human hepatocellular carcinoma) labelling KIF5B with purified ab167429 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

Control : PBS only

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167429).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIF5B+KIF5C antibody [EPR10276(B)] - BSA and Azide free (AB232350)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling KIF5B with Purified ab167429 at 1 : 250 dilution (0.07 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167429).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-KIF5B+KIF5C antibody [EPR10276(B)] - BSA and Azide free (AB232350)

Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling KIF5B with purified ab167429 at 1/20 dilution (10 μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167429).

• IP

Lab

Immunoprecipitation - Anti-KIF5B+KIF5C antibody [EPR10276(B)] - BSA and Azide free (AB232350)

ab167429 (purified) at 1 : 20 dilution (2μg) immunoprecipitating KIF5B in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.

Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab167429 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab167429 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167429).

All lanes:

Immunoprecipitation - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (<a href='/en-us/products/primary-antibodies/kif5bkif5c-antibody-epr10276b-ab167429'>ab167429</a>)

Predicted band size: 110 kDa

false

• WB

Supplier Data

Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] - BSA and Azide free (AB232350)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : KIF5B knockout HAP1 whole cell lysate (20 μg)

Lanes 1 - 2 : Merged signal (red and green). Green - ab167429 observed at 110 kDa. Red - loading control, ab9484, observed at 37 kDa.

Unpurified ab167429 was shown to specifically react with KIF5B in wild-type cells as signal was lost in KIF5B knockout cells. Wild-type and KIF5B knockout samples were subjected to SDS-PAGE. ab167429 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167429).

All lanes:

Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (<a href='/en-us/products/primary-antibodies/kif5bkif5c-antibody-epr10276b-ab167429'>ab167429</a>)

Predicted band size: 110 kDa

false

• WB

Lab

Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] - BSA and Azide free (AB232350)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167429).

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

ab181602 was used a GAPDH loading control.

All lanes:

Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (<a href='/en-us/products/primary-antibodies/kif5bkif5c-antibody-epr10276b-ab167429'>ab167429</a>) at 1/1000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) +OE-empty at 20 µg

Lane 2:

293T (Human embryonic kidney epithelial cell) +OE-mouse KIF5A at 20 µg

Lane 3:

293T (Human embryonic kidney epithelial cell) +OE-mouse KIF5C at 20 µg

Lane 4:

293T (Human embryonic kidney epithelial cell) +OE-mouse KIF5B at 20 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 1/20000 dilution

Observed band size: 110 kDa

false

Exposure time: 1s