Recombinant Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22397-264] to N Cadherin - BSA and Azide free
- Suitable for: WB, Flow Cyt, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free
See all N Cadherin primary antibodies -
Description
Rabbit monoclonal [EPR22397-264] to N Cadherin - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt, IPmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: HeLa, PC-3, C6, A549, HEK-293T and HepG2 whole cell lysate. Human brain lysate. Mouse brain and heart lysate. Rat brain, heart and liver lysate. Flow Cyt: MCF7 cells. IP: N Cadherin IP in HeLa whole cell lysate.
-
General notes
ab245827 is the carrier-free version of ab245117.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22397-264 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
KO cell lines
-
KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab245827 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
WB |
Use at an assay dependent concentration. Detects a band of approximately 130, 110 kDa (predicted molecular weight: 100 kDa).
|
|
Flow Cyt |
Use at an assay dependent concentration.
|
|
IP |
Use at an assay dependent concentration.
|
Notes |
---|
WB
Use at an assay dependent concentration. Detects a band of approximately 130, 110 kDa (predicted molecular weight: 100 kDa). |
Flow Cyt
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Target
-
Function
Cadherins are calcium dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells; cadherins may thus contribute to the sorting of heterogeneous cell types. CDH2 may be involved in neuronal recognition mechanism. In hippocampal neurons, may regulate dendritic spine density. -
Sequence similarities
Contains 5 cadherin domains. -
Cellular localization
Cell membrane. - Information by UniProt
-
Database links
- Entrez Gene: 1000 Human
- Entrez Gene: 12558 Mouse
- Entrez Gene: 83501 Rat
- Omim: 114020 Human
- SwissProt: P19022 Human
- SwissProt: P15116 Mouse
- SwissProt: Q9Z1Y3 Rat
- Unigene: 464829 Human
see all -
Alternative names
- CADH2_HUMAN antibody
- Cadherin 2 antibody
- Cadherin 2 N cadherin neuronal antibody
see all
Images
-
All lanes : Anti-N Cadherin antibody [EPR22397-264] (ab245117) at 1000 µg
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : cdh2 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 100 kDa
Observed band size: 125 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab245117).
False colour image of Western blot: Anti-N Cadherin antibody [EPR22397-264] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab245117 was shown to bind specifically to N Cadherin. A band was observed at 125 kDa in wild-type HeLa cell lysates with no signal observed at this size in cdh2 knockout cell line ab274934 (knockout cell lysate ab274992).
To generate this image, wild-type and cdh2 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
-
All lanes : Anti-N Cadherin antibody [EPR22397-264] (ab245117) at 1/100000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : CDH2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 100 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab245117).
Lanes 1 - 2: Merged signal (red and green). Green - ab245117 observed at 125 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab245117 was shown to react with N Cadherin in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255377 (knockout cell lysate ab263843) was used. Wild-type and N Cadherin knockout samples were subjected to SDS-PAGE. ab245117 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
-
Flow cytometric analysis of MCF7 (Human breast adenocarcinoma epithelial cell, Left) / HeLa (Human cervix adenocarcinoma epithelial cell, Right) cell lines labeling N Cadherin with ab245117 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) 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.
Negative control: MCF7 (PMID: 9177902).
Gated on viable cells.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245117). -
N Cadherin was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell line) whole cell lysate with ab245117 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245117 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: HeLa whole cell lysate 10 µg (Input).
Lane 2: ab245117 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245117 in HeLa whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 seconds.The molecular weight is consistent with literature (PMID: 8230319).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245117).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
Datasheet download
Certificate of Compliance
References (0)
ab245827 has not yet been referenced specifically in any publications.