Antibodies, Proteins, Kits and Reagents for Life Science

307 566 ₸ Product size

100 µl 307 566 ₸ 10 µl 90 460 ₸

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Anti-COL18A1 antibody [EPR23481-240] (ab275390)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23481-240] to COL18A1
Suitable for: Flow Cyt, WB, ICC, IP
Reacts with: Human

Product name

Anti-COL18A1 antibody [EPR23481-240]
See all COL18A1 primary antibodies
Description

Rabbit monoclonal [EPR23481-240] to COL18A1
Host species

Rabbit
Specificity

This antibody does not recognise Endostatin.

This antibody ab275390 [EPR23481-240]] is a distinct clone from ab275746 [EPR23481-21].

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human lung, kidney, placenta and testis tissue lysates; HepG2 and Huh7 whole cell lysates. ICC: HepG2 cells. Flow cyt: HepG2 cells. IP: Huh7 whole cell lysate; Human lung tissue lysate.
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

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23481-240
Isotype

IgG

Immunocytochemistry - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling COL18A1 with ab275390 at 1/50 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 HepG2 cells. Negative control: U937 cell (PMID: 15451191). 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 Fluo^r® 488) at 1/1000 dilution.
Flow Cytometry - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized U-937 (human histiocytic lymphoma monocyte, Left) / HepG2 (human hepatocellular carcinoma epithelial cell, Right) cells labelling COL18A1 with ab275390 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) 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.

Negative control: U937 cell (PMID: 15451191).
Western blot - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

All lanes : Anti-COL18A1 antibody [EPR23481-240] (ab275390) at 1/1000 dilution

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2 : Huh7 (human hepatocellar carcinoma epithelial cell), whole cell lysate at 40 µg
Lane 3 : U937 (human histiocytic lymphoma monocyte), whole cell lysate at 40 µg

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

Predicted band size: 178 kDa
Observed band size: 178,23,28 kDa
why is the actual band size different from the predicted?

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

The expression profile observed is consistent with what has been described in the literature (PMID: 11581192, 16465514, 15451191).

Negative control: U937 (PMID:15451191).

The lanes 1 of blot were developed using a higher sensitivity ECL substrate.

Exposure time: 92 seconds.
Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

COL18A1 was immunoprecipitated from 0.35 mg Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab275390 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275390 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug

Lane 2: ab275390 IP in Huh7 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275390 in Huh7 whole cell lysate

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

Exposure time: Panel A: 32 seconds
Panel B: 3 minutes

Western blot was performed with the immuno-precipitate of ab275390, probed with ab275390 at 1/1000 dilution (Panel A) or ab207162 at 1/1000 dilution (Panel B). The 23- and 28-kDa fragments were recognized by ab207126, suggesting that these fragments are part of the C-terminal non-collagenous domain containing endostatin.
Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

COL18A1 was immunoprecipitated from 0.35 mg Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab275390 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275390 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Huh7 (human hepatocellular carcinoma epithelial cell), whole cell lysate 10 ug

Lane 2: ab275390 IP in Huh7 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275390 in Huh7 whole cell lysate

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

Exposure time: 101 seconds.

23 kDa and 28 kDa fragments are part of the C-terminal non-collagenous domain containing endostatin.
Western blot - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

All lanes : Anti-COL18A1 antibody [EPR23481-240] (ab275390) at 1/1000 dilution

Lane 1 : Human lung tissue lysate
Lane 2 : Human kidney tissue lysate
Lane 3 : Human placenta tissue lysate
Lane 4 : Human testis tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 178 kDa
Observed band size: 23,28 kDa why is the actual band size different from the predicted?

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

The expression profile observed is consistent with what has been described in the literature (PMID: 11581192, 16465514, 15451191).

Exposure time: Lane 1, 4: 3 minutes Lane 2, 3: 26 seconds.
Immunoprecipitation - Anti-COL18A1 antibody [EPR23481-240] (ab275390)

COL18A1 was immunoprecipitated from 0.35 mg human lung tissue lysate with ab275390 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275390 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Human lung tissue lysate 10 ug

Lane 2: ab275390 IP in Human lung tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275390 in Human lung tissue lysate

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

Exposure time: 7.75 seconds.

23 kDa and 28 kDa fragments are part of the C-terminal non-collagenous domain containing endostatin.
Anti-COL18A1 antibody [EPR23481-240] (ab275390)

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