Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸ 10 µl 130 665 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CD166 antibody [EPR2759(2)] (ab109215)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR2759(2)] to CD166
Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-CD166 antibody [EPR2759(2)]
See all CD166 primary antibodies
Description

Rabbit monoclonal [EPR2759(2)] to CD166
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: SH-SY5Y, HuT-78, HT1080, Daudi and HeLa whole cell lysate (ab150035). Mouse and rat brain tissue lysates; Wild-type HAP1 whole cell lysate. IHC-P: Human liver and prostatic adenocarcinoma tissues. ICC/IF: THP-1 cells. Flow Cyt: HuT-78 cells. IP: SH-SY5Y cell 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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR2759(2)
Isotype

IgG
Research areas

Flow Cytometry - Anti-CD166 antibody [EPR2759(2)] (ab109215)

Flow Cytometry analysis of HuT-78 cells labelling CD166 with purified ab109215 at 1/90 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunocytochemistry/ Immunofluorescence - Anti-CD166 antibody [EPR2759(2)] (ab109215)

Immunocytochemistry/Immunofluorescence analysis of THP-1 (human monocytic leukemia cell line) cells labelling CD166 (green) with purified ab109215 at 1/250. Cells were fixed with 100% methanol. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as a nuclear counterstain.

Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD166 with purified ab109215 at 1/50. 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.
Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: CD166 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: SH-SY5Y whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab109215 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab109215 was shown to specifically react with CD166 in wild-type HAP1 cells as signal was lost in CD166 knockout cells. Wild-type and CD166 knockout samples were subjected to SDS-PAGE. Ab109215 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunoprecipitation - Anti-CD166 antibody [EPR2759(2)] (ab109215)

ab109215 (purified) at 1/30 immunoprecipitating CD166 in SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)

All lanes : Anti-CD166 antibody [EPR2759(2)] (ab109215) at 1/10000 dilution (purified)

Lane 1 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate
Lane 2 : HuT-78 cell lysate
Lane 3 : HT-1080 (human fibrosarcoma cell line) cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 65 kDa
Observed band size: 100-105 kDa why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)

All lanes : Anti-CD166 antibody [EPR2759(2)] (ab109215) at 1/10000 dilution (purified)

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

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 65 kDa
Observed band size: 100-105 kDa why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD166 with unpurified ab109215 at 1/100.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)

All lanes : Anti-CD166 antibody [EPR2759(2)] (ab109215) at 1/1000 dilution (unpurified)

Lane 1 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate
Lane 2 : HuT-78 cell lysate
Lane 3 : HT1080 (human fibrosarcoma cell line) cell lysate
Lane 4 : Daudi (human Burkitt's lymphoma cell line) cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 65 kDa
Observed band size: 100-105 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic adenocarcinoma tissue labelling CD166 with unpurified ab109215 at 1/100.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-CD166 antibody [EPR2759(2)] (ab109215)

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