Anti-Collagen I antibody [EPR7785] - Carrier free (ab215969)

Anti-Collagen I [EPR7785] antibody (ab215969) is a rabbit monoclonal antibody that is provided in a PBS only formulation free from BSA and azide. Formulated to be conjugation-ready. It is used to detect Collagen I in Western Blot, IHC-P, IHC-Fr, ICC/IF. Suitable for Human samples.

- Specificity confirmed with Collagen I knockout cell line validation

-BSA,?sodium azide, and glycerol-free?for consistent conjugation with fluorochromes, enzymes and more

Related conjugates and formulations

ab138492
Unconjugated
Anti-Collagen I antibody [EPR7785]
ab313018
Conjugated
Alexa Fluor® 568 Anti-Collagen I antibody [EPR7785]
ab317799
Carrier free
Anti-Collagen I antibody [EPR7785] – Mouse IgG1 (Chimeric) – BSA and Azide Free
ab321114
Conjugated
Alexa Fluor® 750 Anti-Collagen I antibody [EPR7785]
ab280968
Conjugated
Alexa Fluor® 647 Anti-Collagen I antibody [EPR7785]
ab275996
Conjugated
Alexa Fluor® 488 Anti-Collagen I antibody [EPR7785]
ab313221
Conjugated
Alexa Fluor® 555 Anti-Collagen I antibody [EPR7785]
ab311739
Conjugated
Alexa Fluor® 594 Anti-Collagen I antibody [EPR7785]

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (AB215969), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 99% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-Fr	ICC/IF	IHC-P
Human	Tested	Tested	Tested	Tested
Cow	Predicted	Predicted	Predicted	Predicted

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	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. Continuous refrigeration throughout collagen extraction is important to avoid degradation and denaturation. Take care with pH, temperature, and concentration to avoid collagen polymerization. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Predicted
Predicted

Species	Dilution info	Notes
Species Cow	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Cow	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Cow	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab199376 - 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.

Predicted
Predicted

Species	Dilution info	Notes
Species Cow	Dilution info -	Notes -

Associated Products

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4 products for Alternative Version

Target data

See full target information COL1A1

Function

Type I collagen is a member of group I collagen (fibrillar forming collagen).

Alternative names

Collagen alpha-1(I) chain, Alpha-1 type I collagen, COL1A1

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: 99% PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Carrier free

Yes

Clone number

EPR7785

Purification technique

Affinity purification Protein A

Specificity

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.

Dissociation constant

1.22 x 10^-10 M

Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab215969 is the carrier-free version of Anti-Collagen I antibody [EPR7785] ab138492.

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

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.

13 product images

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.
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).
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).
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
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).
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.
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.
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.
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).
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.
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.
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
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.