Anti-Collagen I antibody [EPR7785] - BSA and Azide free

13 product images

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  Image from Kishi Y et al., PLoS One. 2017;12(12):e0189522. Fig 2.; doi: 10.1371/journal.pone.0189522. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  Type I collagen (Anti-Collagen I antibody [EPR7785] ab138492) and Type III collagen immunostainings for control, Subtype I, and Subtype II adenomyotic cases.
  

  The human type I collagen staining bands for adenomyotic cases were thicker than those of the control uteri, and were seen with more fine muscle bundles. Arrowheads indicate vascular walls. Original magnification: X100. Scale bar = 50μm.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Collagen I antibody [EPR7785] ab138492).

  Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunocytochemistry/ Immunofluorescence - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Collagen I antibody [EPR7785] ab138492). Anti-Collagen I antibody [EPR7785] ab138492 staining Collagen alpha-1 chain in wild-type U2OS cells (top panel) and COL1A1 knockout U2OS cells (bottom panel) (Human COL1A1 (PICP) knockout U-2 OS cell line ab273846). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-Collagen I antibody [EPR7785] ab138492 at 0.4μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labelling Collagen I with purified Anti-Collagen I antibody [EPR7785] ab138492 at 1/1500. 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 (Anti-Collagen I antibody [EPR7785] ab138492).

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  Western blot - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  This data was developed using Anti-Collagen I antibody [EPR7785] ab138492, the same antibody clone in a different buffer formulation. Different batches of Anti-Collagen I antibody [EPR7785] ab138492 were tested on HFF-1 (human skin fibroblast) lysate at 0.5 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 220 kDa.

  All lanes: Western blot - Anti-Collagen I antibody [EPR7785] (Anti-Collagen I antibody [EPR7785] ab138492)

  Predicted band size: 139 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labelling Collagen I with unpurified Anti-Collagen I antibody [EPR7785] ab138492 at 1/1500. 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 (Anti-Collagen I antibody [EPR7785] ab138492).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  Immunohistochemistry of breast carcinoma staining Collagen I with Anti-Collagen I antibody [EPR7785] ab138492 at 0.5μg/ml
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Collagen I antibody [EPR7785] ab138492).

  Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  Formalin/PFA-fixed paraffin-embedded sections of human breast carcinoma tissue stained for Collagen I with unpurified Anti-Collagen I antibody [EPR7785] ab138492 in immunohistochemical analysis.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Collagen I antibody [EPR7785] ab138492).

  Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  Formalin/PFA-fixed paraffin-embedded sections of human colon tissue staining Collagen I with unpurified Anti-Collagen I antibody [EPR7785] ab138492 in immunohistochemical analysis.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Collagen I antibody [EPR7785] ab138492).

  Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  IHC image of Collagen I staining in human placenta formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval (EDTA based pH 9.0 solution, epitope retrieval solution 2) for 20 minutes. The section was then incubated with Anti-Collagen I antibody [EPR7785] ab138492 at 1/1500, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Collagen I antibody [EPR7785] ab138492).

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  This image is courtesy of an anonymous customer review.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  Paraffin-embedded human skin tissue stained for Collagen I using Anti-Collagen I antibody [EPR7785] ab138492 at 1/3000 dilution in immunohistochemical analysis, followed by Goat anti rabbit Alexa Fluor­^® 555.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Collagen I antibody [EPR7785] ab138492).

  Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  OI-RD Scanning - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  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.

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  Western blot - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  Blocking buffer and concentration: 5% NFDM/TBST.
  
  Diluting buffer and concentration: 5% NFDM/TBST.
  
  Exposure Time: Lane 1-7: 180 seconds, Lane 8: 60 seconds.
  
  Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as loading control.
  
  Compared with Anti-Collagen I antibody [EPR7785] ab138492, Anti-Collagen I antibody [EPR22209-75] ab255809 has higher affinity. We recommend Anti-Collagen I antibody [EPR22209-75] ab255809 as an alternative for testing pro-Collagen forms in western blot.
  
  Anti-Collagen I antibody [EPR7785] ab138492 is specific for pro-Collagen and 139kda mature from, while Anti-Collagen I antibody [EPR22209-75] ab255809 is specific for pro-Collagen and 35kda C-terminal pro peptide.
  
  For better using Anti-Collagen I antibody [EPR22209-75] ab255809, we recommend loading higher amount of lysate or using lower antibody dilution. 
  
  This data was developed using Anti-Collagen I antibody [EPR7785] ab138492, the same antibody clone in a different buffer formulation.

  All lanes: Western blot - Anti-Collagen I antibody [EPR7785] (Anti-Collagen I antibody [EPR7785] ab138492) at 1/1000 dilution

  Lane 1: A549 (Human lung carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) supernatant lysate at 20 µg

  Lane 4: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 5: SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 6: Human lung tissue lysate at 20 µg

  Lane 7: Human liver tissue lysate at 20 µg

  Lane 8: HFF-1 (Human Skin fibroblast) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 139 kDa

  Observed band size: 220 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (ab215969)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Collagen I antibody [EPR7785] ab138492).

  Immunohistochemical analysis of formalin fixed paraffin embedded human colon cancer labelling Collagen I with Anti-Collagen I antibody [EPR7785] ab138492 at a concentration of 0.89 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit followed by OptiView Amplification kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). Anti-Collagen I antibody [EPR7785] ab138492 anti-Collagen I antibody [EPR7785] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.