Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

ab76126 showing positive staining in human colonic adenocarcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

ab76126 showing negative staining in human hepatocellular carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

Overlay histogram showing LoVo (Human colorectal adenocarcinoma cell line) cells stained with ab76126 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab76126, 1/1000 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 (ab76126).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

ab76126 showing positive staining in human urinary bladder transitional carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma) cells labelling Cytokeratin 20 with purified ab76126 at 1/500. Cells were fixed with 100% methanol. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

Secondary Only Control : PBS was used instead of the primary antibody as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

ab76126 showing negative staining in human breast carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

Fluorescent immunohistochemical analysis of paraffin-embedded human colonic adenocarcinoma tissue using ab76126. Green-CK20 red-PI

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

ab76126 at 1/100 dilution staining Cytokeratin 20 in human colon adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

False colour image of Western blot : Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab76126 was shown to bind specifically to Cytokeratin 20. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).

All lanes:

Western blot - Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker (<a href='/en-us/products/primary-antibodies/cytokeratin-20-antibody-epr1622y-cytoskeleton-marker-ab76126'>ab76126</a>) at 1/10000 dilution

Lane 1:

HT-29 cell lysate at 20 µg

Lane 2:

DLD-1 cell lysate at 20 µg

Lane 3:

SW480 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Lane 5:

MCF7 cell lysate at 20 µg

Predicted band size: 48 kDa

Observed band size: 45 kDa,45-50 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (AB219589)

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.