Anti-COL1A2 antibody [EPR27415-30] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-COL1A2 antibody [EPR27415-30] - BSA and Azide free (AB308456)

This data was developed using ab308455, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell, Left) / WI-38 (human fetal lung fibroblast, Right) cells labelling COL1A2 with ab308455 at 1/50 dilution (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 : HeLa.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COL1A2 antibody [EPR27415-30] - BSA and Azide free (AB308456)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labelling COL1A2 with ab308455 at 1/10000 (0.05 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on interstitial cells in human breast. The section was incubated with ab308455 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COL1A2 antibody [EPR27415-30] - BSA and Azide free (AB308456)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labelling COL1A2 with ab308455 at 1/10000 (0.05 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on interstitial cells in human colon cancer. The section was incubated with ab308455 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COL1A2 antibody [EPR27415-30] - BSA and Azide free (AB308456)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling COL1A2 with ab308455 at 1/10000 (0.05 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on interstitial cells in human colon. The section was incubated with ab308455 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-COL1A2 antibody [EPR27415-30] - BSA and Azide free (AB308456)

This data was developed using ab308455, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized WI-38 (human fetal lung fibroblast) cells labelling COL1A2 with ab308455 at 1/50 (10.06 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in WI-38 cell line, and no staining was observed in HeLa cell line.Negative control : HeLa. 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 2.5ug/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 2ug/ml dilution.

• WB

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Western blot - Anti-COL1A2 antibody [EPR27415-30] - BSA and Azide free (AB308456)

This data was produced using ab308455, the same antibody clone but in a different formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : HeLa. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 15 seconds.

All lanes:

Western blot - Anti-COL1A2 antibody [EPR27415-30] (<a href='/en-us/products/primary-antibodies/col1a2-antibody-epr27415-30-ab308455'>ab308455</a>) at 1/1000 dilution

Lane 1:

WI-38 (human fetal lung fibroblast) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell) 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: 129 kDa

Observed band size: 160 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-COL1A2 antibody [EPR27415-30] - BSA and Azide free (AB308456)

This data was produced using ab308455, the same antibody clone but in a different formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 73 seconds.

All lanes:

Western blot - Anti-COL1A2 antibody [EPR27415-30] (<a href='/en-us/products/primary-antibodies/col1a2-antibody-epr27415-30-ab308455'>ab308455</a>) at 1/1000 dilution

Lane 1:

WI-38 (human fetal lung fibroblast) whole cell lysate at 20 µg

Lane 2:

WI-38 cells transfected with siRNA specifically targeting COL1A2 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: 129 kDa

Observed band size: 160 kDa

false

Exposure time: 73s

• Dot

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Dot Blot - Anti-COL1A2 antibody [EPR27415-30] - BSA and Azide free (AB308456)

This data was developed using ab308455, the same antibody clone in a different buffer formulation. Dot blot analysis of COL1A2 using ab308455 at 1 : 1000 (0.503 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Exposure time : 3 minutes Blocking and diluting buffer and concentration : 5% NFDM/TBST