Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-Collagen I antibody [EPR24331-53] - BSA and Azide free
See all Collagen I primary antibodies
Description

Rabbit monoclonal [EPR24331-53] to Collagen I - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC	Mouse
IHC-Fr	Rat
IHC-P	Mouse
IP	Mouse
WB	Mouse

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: NIH/3T3 whole cell lysate; Mouse skin and Rat skin tissue lysates. IHC-P: Mouse skin, stomach and pancreatic cancer tissue; Rat skin tissue. IHC-Fr: Rat skin; Mouse skin tissue. Flow Cyt (intra): NIH/3T3 cells. ICC: NIH/3T3 cells. IP: Mouse skin tissue lysate.
General notes
ab279711 is the carrier-free version of ab270993. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab279711 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.

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

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C.
Storage buffer

Constituent: 100% PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24331-53
Isotype

IgG
Research areas

Western blot - Anti-Collagen I antibody [EPR24331-53] - BSA and Azide free (ab279711)

All lanes : Anti-Collagen I antibody [EPR24331-53] (ab270993) at 1/1000 dilution

Lane 1 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg
Lane 2 : NIH/3T3 culture supernatant at 20 µl

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 139 kDa

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Cleaved C-propeptide (30-40kD) and pCcollagen (75-100kD) are observed. The molecular weight observed is consistent with what has been described in the literature (PMID: 23940311;PMID: 29853175).

Exposure time: 92 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - (ab279711)

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse skin tissue labeling Collagen I with ab270993 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in connective tissues of mouse skin. The section was incubated with ab270993 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - (ab279711)

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling Collagen I with ab270993 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in connective tissues of mouse stomach. The section was incubated with ab270993 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - (ab279711)

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse pancreatic cancer tissue labeling Collagen I with ab270993 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in connective tissues of mouse pancreatic cancer. The section was incubated with ab270993 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Frozen sections) - (ab279711)

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skin tissue labeling Collagen I with ab270993 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse skin is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunocytochemistry - (ab279711)

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling Collagen I with ab270993 at 1/2000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Flow Cytometry - (ab279711)

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling Collagen I with ab270993 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - (ab279711)

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Collagen I was immunoprecipitated from 0.35 mg Mouse skin tissue lysate 10 ug with ab270993 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270993 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse skin tissue lysate 10 ug

Lane 2: ab270993 IP in Mouse skin tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab270993 in Mouse skin tissue lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 32 seconds
Western blot - Anti-Collagen I antibody [EPR24331-53] - BSA and Azide free (ab279711)

All lanes : Anti-Collagen I antibody [EPR24331-53] (ab270993) at 1/1000 dilution

Lane 1 : Mouse skin tissue lysate
Lane 2 : Rat skin tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 139 kDa
Observed band size: 138 kDa why is the actual band size different from the predicted?

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST

The observed MW is consistent with what has been described in the literature (PMID: 27740527;PMID: 22278938; PMID: 26973392).

Exposure time: Lane 1: 3.25 seconds

Lane 2: 5.5 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - (ab279711)

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat skin tissue labeling Collagen I with ab270993 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in connective tissues of rat skin. The section was incubated with ab270993 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Frozen sections) - (ab279711)

This data was developed using ab270993, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat skin tissue labeling Collagen I with ab270993 at 1/500 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on rat skin is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).