Anti-Collagen II antibody [EPR27418-25]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen II antibody [EPR27418-25] (AB307674)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling Collagen II with ab307674 at 1/200 (2.34 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on rat skeletal muscle.The section was incubated with ab307674 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Collagen II antibody [EPR27418-25] (AB307674)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh) tissue labeling Collagen II with ab307674 at 1/100 (4.68 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control : confocal image showing no staining on rat liver (PMID 9121483). The nuclear counterstain was DAPI (Blue). The section was incubated with ab307674 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen II antibody [EPR27418-25] (AB307674)

Immunohistochemical analysis of paraffin-embedded Rat cartilage tissue labeling Collagen II with ab307674 at 1/200 (2.34 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cartilage.The section was incubated with ab307674 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Collagen II antibody [EPR27418-25] (AB307674)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat articular cartilage (fresh) tissue labeling Collagen II with ab307674 at 1/100 (4.68 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on rat articular cartilage. The nuclear counterstain was DAPI (Blue). The section was incubated with ab307674 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• WB

Supplier Data

Western blot - Anti-Collagen II antibody [EPR27418-25] (AB307674)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : skeletal muscle (Human Protein Atlas Database), liver (PMID : 9121483). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Collagen II antibody [EPR27418-25] (ab307674) at 1/1000 dilution

Lane 1:

Human articar cartilage tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 142 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Collagen II antibody [EPR27418-25] (AB307674)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The 142kDa and 110kDa bands represent full length of Collagen II and Collagen alpha-1(II) chain. Exposure time : 26 seconds

All lanes:

Western blot - Anti-Collagen II antibody [EPR27418-25] (ab307674) at 1/1000 dilution

All lanes:

Rat articar cartilage tissue 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 1/20000 dilution

Observed band size: 110 kDa,142 kDa

false

Exposure time: 26s