Anti-5T4 antibody [EPR5529] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue labelling 5T4 with purified ab134162 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134162).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711)

Immunohistochemical staining using ab134162 on paraffin embedded human placenta tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134162).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue labelling 5T4 with unpurified ab134162 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134162).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711)

Immunohistochemical staining using ab134162 on paraffin embedded human endometrial cancer.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134162).

• WB

Unknown

Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711)

This data was developed using ab134162, the same antibody clone in a different buffer formulation :

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711) at 1/5000 dilution

All lanes:

HT-29 cell lysate at 20 µg

Secondary

All lanes:

Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 46 kDa

Observed band size: 72-80 kDa

false

• WB

Unknown

Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711)

This data was developed using ab134162, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711) at 1/500 dilution

All lanes:

HT-29 cell lysate at 20 µg

Secondary

All lanes:

Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 46 kDa

Observed band size: 72-80 kDa

false

• WB

Lab

Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711)

This data was developed using the same antibody clone in a different buffer formulation (ab134162).

Lanes 1 - 4 : Merged signal (red and green). Green - ab134162 observed at 85 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab134162 was shown to react with 5T4 in wild-type MCF7 cells in Western blot with loss of signal observed in TPBG knockout cell line ab269499 (knockout cell lysate ab269661). Wild-type MCF7 and TPBG knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in before incubation with ab134162 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-5T4 antibody [EPR5529] (<a href='/en-us/products/primary-antibodies/5t4-antibody-epr5529-ab134162'>ab134162</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

TPBG knockout MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TPBG knockout MCF7 cell line (<a href='/en-us/products/cell-lines/human-tpbg-knockout-mcf7-cell-line-ab269499'>ab269499</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 85 kDa

false

• WB

Unknown

Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711)

This data was developed using ab134162, the same antibody clone in a different buffer formulation

All lanes:

Western blot - Anti-5T4 antibody [EPR5529] (<a href='/en-us/products/primary-antibodies/5t4-antibody-epr5529-ab134162'>ab134162</a>) at 1/1000 dilution

Lane 1:

HT-29 cell lysates at 10 µg

Lane 2:

HT-1376 cell lysates at 10 µg

Lane 3:

Caco-2 cell lysates at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 46 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.