Anti-Collagen I antibody [EPR7785] ab138492 is a rabbit monoclonal antibody that is used in Collagen I western blotting, IHC and immunofluorescence. Suitable for human samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with COL1A1 knockout cell line validation
- Antibody clone EPR7785 is cited in over 350 publications
IgG
Rabbit
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
ICC/IF | WB | IHC-Fr | IHC-P | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Cow | Predicted | Predicted | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.4 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Cow | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 - 1/10000 | Notes Sample preparation: frozen, ground tissue samples were added to hot lysis buffer (10 mM Tris-HCl (pH 8.0); 1%SDS; 1.0 mM Na-Orthovanadate), boiled for 10-20 minutes and sonicated (3 seconds at 40kW, 30 intervals) prior to centrifugation. Positive Control: Hu stomach, skin and adrenal gland tissue lysates. Acid or enzyme treatment with pepsin is a better method to isolate collagen. |
Species | Dilution info | Notes |
---|---|---|
Species Cow | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.1-0.5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Cow | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1500 | Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Cow | Dilution info - | Notes - |
Select an associated product type
Type I collagen is a member of group I collagen (fibrillar forming collagen).
Collagen alpha-1(I) chain, Alpha-1 type I collagen, COL1A1
Anti-Collagen I antibody [EPR7785] ab138492 is a rabbit monoclonal antibody that is used in Collagen I western blotting, IHC and immunofluorescence. Suitable for human samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with COL1A1 knockout cell line validation
- Antibody clone EPR7785 is cited in over 350 publications
IgG
Rabbit
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR7785
Affinity purification Protein A
Compared with ab138492, ab255809 has higher affinity. We recommend ab255809 as an alternative for testing pro-Collagen forms in western blot. ab138492 works in western blot in samples with high level of collagen I, like HFF-1, MRC-5, skin tissue etc.
ab138492 is specific for pro-Collagen and 139kda mature from, while ab255809 is specific for pro-Collagen and 35kda C-terminal pro peptide.
Please navigate to the support & downloads section for specificity details in Chinese.
1.22 x 10-10 M
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
Collagen type I also called collagen I is a structural protein expressed mainly in connective tissues such as skin tendon bone and ligaments. It serves as an important component in providing mechanical strength and integrity to these tissues. Collagen I is a fibrillar collagen known for its triple-helix structure composed of two alpha-1 chains and one alpha-2 chain and has a molecular mass of approximately 300 kDa. Researchers often employ collagen western blot and collagen ELISA techniques for its detection. Collagen suppliers offer various collagen antibodies used in these assays to study its distribution and function.
Collagen type I plays a central role in maintaining the extracellular matrix and supporting cellular environments. It interacts with other matrix proteins and cells forming complexes that help in tissue development and repair. Type I collagen is especially important in bone matrix working alongside minerals like hydroxyapatite to provide rigidity and support. Anti-collagen antibodies aid in studying its biological functions and interactions which are critical to understanding tissue dynamics.
Collagen type I interacts with multiple signaling cascades involved in tissue remodeling and repair. It is a significant player in the TGF-β pathway which regulates fibrosis and wound healing processes. In these pathways proteins such as fibronectin and integrins work in concert with collagen type I to orchestrate cellular responses to damage. Researchers often examine its role in these pathways to uncover therapeutic possibilities for disease interventions.
Collagen type I has strong connections to conditions like osteogenesis imperfecta and fibrosis. Mutations or irregularities in collagen I production can lead to osteogenesis imperfecta a genetic disorder characterized by brittle bones. In fibrosis excessive collagen deposition disrupts normal tissue architecture contributing to organ dysfunction. In both conditions type I collagen interacts with other proteins like matrix metalloproteinases which modulate its breakdown and remodeling highlighting its importance in disease pathology.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Western blot results of Collagen I from ADSCs (human adipose derived stem cells) cultured in RAD16-I alone, RAD/CS or RAD/Decorin. Actin was used as an internal control. Samples were prepared in triplicate; control, control medium; chondro, chondrogenic medium.
Samples were lysed in RIPA buffer with a protease inhibitor cocktail. Acrylamide gels were prepared according to the size of the proteins, generally at concentrations of 7.5% or 10% (w/v). Cell lysates (5 mg) were run by applying 150 V for 90 min. Proteins were transferred to a PVDF membrane by applying 40 V for 2 hours at RT. The membrane was incubated at RT for 2 hours in blocking buffer (BB) consisting of 4% (w/v) nonfat milk powder in PBST. Membranes were incubated for 1 hour at RT with ab138492 at a final concentration of 1 mg/mL in PBST. An anti-rabbit (IgG-HRP) secondary antibody was added, at a final concentration of 1 mg/mL, and incubated at RT for 1 h.
For full image please see paper.
All lanes: Western blot - Anti-Collagen I antibody [EPR7785] (ab138492)
Predicted band size: 139 kDa
Type I collagen (ab138492) and Type III collagen immunostainings for control, Subtype I, and Subtype II adenomyotic cases.
The 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.
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 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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labeling Collagen I with purified 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.
Blocking buffer: 5% NFDM/TBST.
Exposure time: 180 seconds.
All lanes: Western blot - Anti-Collagen I antibody [EPR7785] (ab138492) at 1/5000 dilution
Lane 1: HFF-1 (human skin fibroblast) whole cell lysate at 15 µg
Lane 2: MRC-5 (Human lung fibroblast) whole cell lysate at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 139 kDa
Observed band size: 220 kDa
The lysate in this image is prepared by 1%SDS Hot lysis method. For Lysate preparation protocol, please refer to the protocol section of the website and/or here.
All lanes: Western blot - Anti-Collagen I antibody [EPR7785] (ab138492) at 1/1000 dilution
Lane 1: Human stomach tissue lysate at 10 µg
Lane 2: Human skin lysate at 10 µg
Lane 3: Human adrenal gland lysate at 10 µg
Lane 1: HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Lanes 2 - 3: HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 139 kDa
IHC image of Collagen I staining in a section of frozen human cervix* performed on a Leica Biosystems BOND® RX instrumen using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab138492, 0.01 ug/ml, for 15 mins 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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
IHC image of Collagen I staining in a section of frozen human uterus* performed on a Leica Biosystems BOND® RX instrumen using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab138492, 0.05 ug/ml, for 15 mins 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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
Different batches of 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] (ab138492)
Predicted band size: 139 kDa
Frozen, ground tissue samples were added to hot lysis buffer (10 mM Tris-HCl (pH 8.0); 1%SDS; 1.0 mM Na-Orthovanadate), boiled for 10-20 minutes and sonicated (3 seconds, 30 intervals).
The blocking and antibody incubations were performed in 5% non-fat milk (TBST).
The lysate in this image is prepared by 1%SDS Hot lysis method. For Lysate preparation protocol, please refer to the protocol section of the website and/or here.
All lanes: Western blot - Anti-Collagen I antibody [EPR7785] (ab138492) at 1/5000 dilution
All lanes: Human skin tissue lysate at 10 µg
All lanes: HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 139 kDa
Observed band size: 139 kDa
Frozen, ground tissue samples were added to hot lysis buffer (10 mM Tris-HCl (pH 8.0); 1%SDS; 1.0 mM Na-Orthovanadate), boiled for 10-20 minutes and sonicated (3 seconds, 30 intervals).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
TBST).
The lysate in this image is prepared by 1%SDS Hot lysis method. For Lysate preparation protocol, please refer to the protocol section of the website and/or here.
All lanes: Western blot - Anti-Collagen I antibody [EPR7785] (ab138492) at 1/5000 dilution
All lanes: Human skin tissue lysate at 10 µg
All lanes: HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/5000 dilution
Predicted band size: 139 kDa
Observed band size: 139 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labelling Collagen I with unpurified 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.
Immunohistochemistry of human breast carcinoma tissue staining Collagen I with ab138492 at 0.5μg/ml.
Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.
Formalin/PFA-fixed paraffin-embedded sections of human breast carcinoma tissue stained for Collagen I with unpurified ab138492 in immunohistochemical analysis.
Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.
Formalin/PFA-fixed paraffin-embedded sections of human colon tissue staining Collagen I with unpurified ab138492 in immunohistochemical analysis.
Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.
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 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
Paraffin-embedded human skin tissue stained for Collagen I using ab138492 at 1/3000 dilution in immunohistochemical analysis, followed by Goat anti rabbit Alexa Fluor® 555.
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.
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 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.
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.
All lanes: Western blot - 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
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
Immunohistochemical analysis of formalin fixed paraffin embedded human colon cancer labelling Collagen I with 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). 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.
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