Anti-Cdc34 antibody [EPR16808] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Cdc34 antibody [EPR16808] - BSA and Azide free (AB251415)

This data was developed using ab204515, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Cdc34 with ab204515 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cdc34 antibody [EPR16808] - BSA and Azide free (AB251415)

This data was developed using ab204515, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Cdc34 with ab204515 at 1/50 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows :
-ve control 1 : ab204515 at 1/50 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Cdc34 antibody [EPR16808] - BSA and Azide free (AB251415)

This data was developed using ab204515, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Cdc34 with ab204515 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-Cdc34 antibody [EPR16808] - BSA and Azide free (AB251415)

This data was developed using ab204515, the same antibody clone in a different buffer formulation.

Cdc34 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab204515 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab204515 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : K562 whole cell lysate 10ug (Input).

Lane 2 : ab204515 IP in K562 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab204515 in K562 whole cell lysate.

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

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-Cdc34 antibody [EPR16808] (<a href='/en-us/products/primary-antibodies/cdc34-antibody-epr16808-ab204515'>ab204515</a>)

Predicted band size: 26 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cdc34 antibody [EPR16808] - BSA and Azide free (AB251415)

This data was developed using ab204515, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cells) cells labeling Cdc34 with ab204515 at 1/50 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasmic staining on Neuro-2a cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows :
-ve control 1 : ab204515 at 1/50 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-Cdc34 antibody [EPR16808] - BSA and Azide free (AB251415)

This data was developed using ab204515, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Anti-BTK (phospho Y223) antibody [EP420Y] (Alexa Fluor® 488) (ab204516) at 1/5000 dilution

Lane 1:

293 (Human epithelial cells from embryonic kidney) whole cell lysate at 10 µg

Lane 2:

K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate at 10 µg

Lane 3:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/100000 dilution

Predicted band size: 26 kDa

Observed band size: 35 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-Cdc34 antibody [EPR16808] - BSA and Azide free (AB251415)

This data was developed using ab204515, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Cdc34 antibody [EPR16808] (<a href='/en-us/products/primary-antibodies/cdc34-antibody-epr16808-ab204515'>ab204515</a>) at 1/5000 dilution

Lane 1:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 2:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 4:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 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 1/50000 dilution

Predicted band size: 26 kDa

Observed band size: 35 kDa

false

Exposure time: 2s

• WB

Supplier Data

Western blot - Anti-Cdc34 antibody [EPR16808] - BSA and Azide free (AB251415)

This data was developed using ab204515, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Cdc34 antibody [EPR16808] (<a href='/en-us/products/primary-antibodies/cdc34-antibody-epr16808-ab204515'>ab204515</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

Mouse heart tissue lysate at 10 µg

Lane 3:

Mouse kidney tissue lysate at 10 µg

Lane 4:

Rat heart tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 26 kDa

Observed band size: 35 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Cdc34 antibody [EPR16808] - BSA and Azide free (AB251415)

This data was developed using ab204515, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Cdc34 antibody [EPR16808] (<a href='/en-us/products/primary-antibodies/cdc34-antibody-epr16808-ab204515'>ab204515</a>) at 1/5000 dilution

Lane 1:

Recombinant protein fragment human Cdc34 at 0.01 µg

Lane 2:

Recombinant protein fragment human Cdc34B at 0.01 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 1/50000 dilution

Predicted band size: 26 kDa

Observed band size: 35 kDa

false

Exposure time: 1s