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

• IHC-P

Lab

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 (ab138492).

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.

• IHC-P

PubMed

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

Type I collagen (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 (ab138492).

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

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/

• IHC-P

Lab

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 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 (ab138492).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (AB215969)

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. This data was developed using the same antibody clone in a different buffer formulation (ab138492).

• IHC-P

AbReview61068****

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 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 (ab138492).

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

This image is courtesy of an anonymous Abreview.

• IHC-P

Unknown

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 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 (ab138492).

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

• IHC-P

Lab

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 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 (ab138492).

• IHC-P

Lab

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 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 (ab138492).

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

• ICC/IF

Lab

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 (ab138492). ab138492 staining Collagen alpha-1 chain in wild-type U2OS cells (top panel) and COL1A1 knockout U2OS cells (bottom panel) (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 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) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (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).

• IHC-P

Unknown

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 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 (ab138492).

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

• IHC-P

Lab

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 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 (ab138492).

• WB

Lab

Western blot - Anti-Collagen I antibody [EPR7785] - BSA and Azide free (AB215969)

This data was developed using ab138492, the same antibody clone in a different buffer formulation. 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] (<a href='/en-us/products/primary-antibodies/collagen-i-antibody-epr7785-ab138492'>ab138492</a>)

Predicted band size: 139 kDa

false

• WB

Lab

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. ab181602 was used as loading control. Compared with ab138492, ab255809 has higher affinity. We recommend ab255809 as an alternative for testing pro-Collagen forms in western blot. ab138492 is specific for pro-Collagen and 139kda mature from, while ab255809 is specific for pro-Collagen and 35kda C-terminal pro peptide. For better using ab255809, we recommend loading higher amount of lysate or using lower antibody dilution.  This data was developed using ab138492, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Collagen I antibody [EPR7785] (<a href='/en-us/products/primary-antibodies/collagen-i-antibody-epr7785-ab138492'>ab138492</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 139 kDa

Observed band size: 220 kDa

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• OI-RD Scanning

Unknown

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.