Anti-Collagen I 抗体 [EPR7785]

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Collagen I antibody [EPR7785] (AB138492)

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-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] (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

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] (AB138492)

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-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] (AB138492)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] (AB138492)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Collagen I antibody [EPR7785] (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] (AB138492)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] (AB138492)

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.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] (AB138492)

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.

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] (AB138492)

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

• IHC-P

AbReview61068****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen I antibody [EPR7785] (AB138492)

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 image is courtesy of an anonymous Abreview.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Collagen I antibody [EPR7785] (AB138492)

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.

• WB

Lab

Western blot - Anti-Collagen I antibody [EPR7785] (AB138492)

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 139 kDa

Observed band size: 220 kDa

false

• WB

Lab

Western blot - Anti-Collagen I antibody [EPR7785] (AB138492)

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

false

• WB

Lab

Western blot - Anti-Collagen I antibody [EPR7785] (AB138492)

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

Secondary

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

false

• WB

PubMed

Western blot - Anti-Collagen I antibody [EPR7785] (AB138492)

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

false

Recha-Sancho et al PLoS One. 2016 Jun 17;11(6):e0157603. doi: 10.1371/journal.pone.0157603. eCollection 2016. Fig 7. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• WB

Unknown

Western blot - Anti-Collagen I antibody [EPR7785] (AB138492)

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

Secondary

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

false

• WB

Lab

Western blot - Anti-Collagen I antibody [EPR7785] (AB138492)

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

Secondary

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

false

• WB

Lab

Western blot - Anti-Collagen I antibody [EPR7785] (AB138492)

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.

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/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

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Collagen I antibody [EPR7785] (AB138492)

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.