Anti-Collagen IV antibody [EPR20966] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen IV antibody [EPR20966] - BSA and Azide free (AB226485)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Collagen IV with ab214417 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Basement membrane staining on human kidney (PMID : 20551626, PMID : 24445937). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214417).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen IV antibody [EPR20966] - BSA and Azide free (AB226485)

Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue labeling Collagen IV with ab214417 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Basement membrane staining on human thyroid cancer. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214417).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Collagen IV antibody [EPR20966] - BSA and Azide free (AB226485)

False colour image of Western blot : Anti-Collagen IV antibody [EPR20966] staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab214417 was shown to bind specifically to Collagen IV. A band was observed at 200 kDa in wild-type HeLa cell lysates with no signal observed at this size in Col4a1 knockout cell line ab255379 (knockout cell lysate ab263755). To generate this image, wild-type and Col4a1 knockout HeLa cell lysates were analysed. 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.

All lanes:

Western blot - Anti-Collagen IV antibody [EPR20966] (<a href='/en-us/products/primary-antibodies/collagen-iv-antibody-epr20966-ab214417'>ab214417</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Col4a1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-col4a1-collagen-iv-alpha-1-knockout-hela-cell-line-ab255379'>ab255379</a>)

Predicted band size: 161 kDa

Observed band size: 200 kDa

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