Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸ 10 µl 110 563 ₸

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Anti-Collagen I antibody [EPR22894-89] (ab260043)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22894-89] to Collagen I
Suitable for: WB, ICC/IF, Flow Cyt, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-Collagen I antibody [EPR22894-89]
See all Collagen I primary antibodies
Description

Rabbit monoclonal [EPR22894-89] to Collagen I
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse Human
ICC/IF	Mouse Human
IP	Mouse
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- ICC/IF: HFF-1 and NIH/3T3 cells. WB: HFF-1 and NIH/3T3 whole cell lysate. Rat lung organ culture supernatant. Human lung tissue lysate. IP: Mouse lung lysate. Flow Cyt: HFF-1 and NIH/3T3 cells.
General notes
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22894-89
Isotype

IgG
Research areas

Immunocytochemistry/ Immunofluorescence - Anti-Collagen I antibody [EPR22894-89] (ab260043)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HFF-1 (Human skin fibroblast) cells labeling Collagen I with ab260043 at 1/250 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HFF-1 cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - 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 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Collagen I antibody [EPR22894-89] (ab260043)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labeling Collagen I with ab260043 at 1/250 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - 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 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Western blot - Anti-Collagen I antibody [EPR22894-89] (ab260043)

All lanes : Anti-Collagen I antibody [EPR22894-89] (ab260043) at 1/5000 dilution

Lane 1 : HFF-1 (human skin fibroblast) whole cell lysate at 20 µg
Lane 2 : HFF-1 culture supernatant at 10 µl

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/25000 dilution

Predicted band size: 139 kDa
Observed band size: 220,35 kDa
why is the actual band size different from the predicted?

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

Procollagen I (220 kDa) and cleaved C-propeptide (35 kDa) are observed.

The molecular weight observed is consistent with what has been described in the literature (PMID: 23940311).

Exposure time: Lane 1: 1 second; Lane 2: 3 seconds.
Western blot - Anti-Collagen I antibody [EPR22894-89] (ab260043)

All lanes : Anti-Collagen I antibody [EPR22894-89] (ab260043) at 1/1000 dilution

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) culture supernatant at 10 µl
Lane 2 : Rat lung organ culture supernatant at 10 µl
Lane 3 : Human lung tissue lysate at 10 µg

Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/25000 dilution
Lane 3 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Predicted band size: 139 kDa

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

Cleaved collagen I C-propeptide (35 kDa) is observed.

The molecular weight observed is consistent with what has been described in the literature (PMID: 23940311).

Exposure time: Lanes 1-2: 3 seconds; Lane 3: 136 seconds.
Immunoprecipitation - Anti-Collagen I antibody [EPR22894-89] (ab260043)

Collagen I was immunoprecipitated from 0.35 mg mouse lung lysate 10µg with ab260043 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab260043 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Lane 1: Mouse lung lysate 10µg

Lane 2: ab260043 IP in mouse lung lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab260043 in mouse lung lysate

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

Exposure time: 1 second.
Flow Cytometry - Anti-Collagen I antibody [EPR22894-89] (ab260043)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labeling Collagen I with ab260043 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry - Anti-Collagen I antibody [EPR22894-89] (ab260043)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HFF-1 (Human skin fibroblast) cells labeling Collagen I with ab260043 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Anti-Collagen I antibody [EPR22894-89] (ab260043)

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