Anti-Zic2 antibody [EPR7790] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Zic2 antibody [EPR7790] - BSA and Azide free (AB236069)

Overlay histogram showing SH-SY5Y cells stained with unpurified ab150404 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab150404, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150404).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Zic2 antibody [EPR7790] - BSA and Azide free (AB236069)

Immunocytochemistry analysis of SK-OV-3 (human ovarian cancer epithelial cell)cells labelling Zic2 with ab150404 at 2.5 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red). Nuclear DNA was labelled with DAPI (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150404).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Zic2 antibody [EPR7790] - BSA and Azide free (AB236069)

Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Zic2 with purified ab150404 at 1/600 dilution (1 μg/ml) (red). Cells were fixed with 80% 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 (ab150404).

• WB

Unknown

Western blot - Anti-Zic2 antibody [EPR7790] - BSA and Azide free (AB236069)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150404).

All lanes:

Western blot - Anti-Zic2 antibody [EPR7790] (<a href='/ko/products/primary-antibodies/zic2-antibody-epr7790-ab150404'>ab150404</a>) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human cerebellum lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 55 kDa

Observed band size: ~63 kDa

false

• WB

Lab

Western blot - Anti-Zic2 antibody [EPR7790] - BSA and Azide free (AB236069)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150404).

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

All lanes:

Western blot - Anti-Zic2 antibody [EPR7790] (<a href='/ko/products/primary-antibodies/zic2-antibody-epr7790-ab150404'>ab150404</a>) at 1/15000 dilution

All lanes:

Human fetal brain lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 63 kDa

false

• WB

Lab

Western blot - Anti-Zic2 antibody [EPR7790] - BSA and Azide free (AB236069)

Exposure time :

Lane1 : 3.25 seconds
Lane2 : 20 seconds
Lane3 : 5.5 seconds
Lane4-5 : 60 seconds

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

ab150404 detects an 70kDa extra band in some cell lines. The expression profile observed in U-87 MG is consistent with the literature (PMID : 11038359). Negative control : U-87 MG (PMID : 11038359)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150404).

All lanes:

Western blot - Anti-Zic2 antibody [EPR7790] (<a href='/ko/products/primary-antibodies/zic2-antibody-epr7790-ab150404'>ab150404</a>) at 1/500 dilution

Lane 1:

Human fetal brain tissue lysate at 20 µg

Lane 2:

PANC-1 (Human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

SK-OV-3 (Human ovarian cancer epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

Mouse embryo tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 56 kDa

false

• WB

Lab

Western blot - Anti-Zic2 antibody [EPR7790] - BSA and Azide free (AB236069)

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

It detects an 70kDa extra band in some cell lines.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150404).

All lanes:

Western blot - Anti-Zic2 antibody [EPR7790] (<a href='/ko/products/primary-antibodies/zic2-antibody-epr7790-ab150404'>ab150404</a>) at 1/2000 dilution

Lane 1:

Human fetal brain tissue lysate at 20 µg

Lane 2:

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

Lane 3:

C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 4:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 25 kDa,55 kDa

Observed band size: 28 kDa,56 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Zic2 antibody [EPR7790] - BSA and Azide free (AB236069)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150404).

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

All lanes:

Western blot - Anti-Zic2 antibody [EPR7790] (<a href='/ko/products/primary-antibodies/zic2-antibody-epr7790-ab150404'>ab150404</a>) at 1/5000 dilution

All lanes:

Mouse brain lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

false

• WB

Lab

Western blot - Anti-Zic2 antibody [EPR7790] - BSA and Azide free (AB236069)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150404).

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

All lanes:

Western blot - Anti-Zic2 antibody [EPR7790] (<a href='/ko/products/primary-antibodies/zic2-antibody-epr7790-ab150404'>ab150404</a>) at 1/20000 dilution

All lanes:

Rat brain lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 63 kDa

false