Anti-HOXC10 antibody [EPR27318-83] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HOXC10 antibody [EPR27318-83] - BSA and Azide free (AB308519)

This data was developed using ab308518, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling HOXC10 with ab308518 at 1/50 (9.02 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in HeLa cell line.Negative control : HepG2. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). 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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-HOXC10 antibody [EPR27318-83] - BSA and Azide free (AB308519)

This data was developed using ab308518, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell, Left) / HeLa (human cervical adenocarcinoma epithelial cell, Right) cells labelling HOXC10 with ab308518 at 1/500 dilution (0.1 ug)/Red (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 1/5000 dilution was used as the secondary antibody. Negative control : HepG2.

• WB

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Western blot - Anti-HOXC10 antibody [EPR27318-83] - BSA and Azide free (AB308519)

This data was developed using 308518, the same antibody clone in a different buffer formulation.  Blocking and diluting buffer and concentration : 5% NFDM/TBST. Additional bands may be due to post-translational modification by ubiquitination (PMID : 12853486).  The Lane 1 was freshly made and used for Western blotting immediately to minimize protein degradation. The lane 2 was developed using a high sensitivity ECL substrate. Exposure time : Lane 1 : 114 seconds, Lane 2 : 125 seconds

All lanes:

Western blot - Anti-HOXC10 antibody [EPR27318-83] (<a href='/en-us/products/primary-antibodies/hoxc10-antibody-epr27318-83-ab308518'>ab308518</a>) at 1/1000 dilution

Lane 1:

T-47D (human ductal breast epithelial tumor epithelial cell), whole cell lysate at 20 µg

Lane 2:

Human breast tissue lysate 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 1/100000 dilution

Predicted band size: 38 kDa

Observed band size: 40 kDa

false

• WB

Supplier Data

Western blot - Anti-HOXC10 antibody [EPR27318-83] - BSA and Azide free (AB308519)

This data was developed using 308518, the same antibody clone in a different buffer formulation.  Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : K562, HL-60 (PMID : 7527557), HepG2 Additional bands may be due to post-translational modification by ubiquitination (PMID : 12853486). This blot was developed using a higher sensitivity ECL substrate. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 169 seconds

All lanes:

Western blot - Anti-HOXC10 antibody [EPR27318-83] (<a href='/en-us/products/primary-antibodies/hoxc10-antibody-epr27318-83-ab308518'>ab308518</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 5:

HL-60 (human Acute Promyelocytic Leukemia promyeloblast) whole cell lysate 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 1/100000 dilution

Predicted band size: 38 kDa

Observed band size: 40 kDa

false

Exposure time: 169s

• WB

Supplier Data

Western blot - Anti-HOXC10 antibody [EPR27318-83] - BSA and Azide free (AB308519)

This data was developed using 308518, the same antibody clone in a different buffer formulation.  Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 3 seconds

All lanes:

Western blot - Anti-HOXC10 antibody [EPR27318-83] (<a href='/en-us/products/primary-antibodies/hoxc10-antibody-epr27318-83-ab308518'>ab308518</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell)cells transfected with an empty vector containing a his tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human HOXC10 expression vector containing a his tag whole cell lysate 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 1/100000 dilution

Predicted band size: 38 kDa

Observed band size: 40 kDa

false

Exposure time: 3s