Anti-Eg5 antibody [EPR23277-60] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] - BSA and Azide free (AB270409)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Eg5 with ab254299 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Cytoplasmic staining in human tonsil (PMID : 25277178) is observed.

The section was incubated with ab254299 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254299).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Eg5 antibody [EPR23277-60] - BSA and Azide free (AB270409)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell line) cells labelling Eg5 with ab254299 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cells. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor^® 594) was used to counterstain tubulin at 1/100 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254299).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] - BSA and Azide free (AB270409)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Eg5 with ab254299 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Cytoplasmic staining in human tonsil (PMID : 25277178) is observed. The section was incubated with ab254299 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254299).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Eg5 antibody [EPR23277-60] - BSA and Azide free (AB270409)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Eg5 with ab254299 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254299).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] - BSA and Azide free (AB270409)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Eg5 with ab254299 at 1/500 dilution (1.18 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Cytoplasmic staining in human tonsil (PMID : 25277178) is observed.

The section was incubated with ab254299 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254299).

• IP

Lab

Immunoprecipitation - Anti-Eg5 antibody [EPR23277-60] - BSA and Azide free (AB270409)

Eg5 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate with ab254299 at 1/30 dilution. Western blot was performed on the immunoprecipitate using 254299 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa whole cell lysate 10 μg

Lane 2 : ab254299 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254299 in HeLa whole cell lysate.

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

Exposure time : 3 seconds

The minor bands beneath the target band are probably caused by degradation.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254299).

All lanes:

Immunoprecipitation - Anti-Eg5 antibody [EPR23277-60] (<a href='/en-us/products/primary-antibodies/eg5-antibody-epr23277-60-ab254299'>ab254299</a>)

Predicted band size: 119 kDa

Observed band size: 130 kDa

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