Mouse Monoclonal I Afadin antibody. Carrier free. Suitable for WB and reacts with Human, Mouse, Rat samples.
Images
![Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (AB300636), expandable thumbnail](https://content.abcam.com/products/images/af6-antibody-35-af6-bsa-and-azide-free-ab300636--western-blot-img404511.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (AB300636)")
![Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (AB300636), expandable thumbnail](https://content.abcam.com/products/images/af6-antibody-35-af6-bsa-and-azide-free-ab300636--western-blot-img404512.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (AB300636)")
Key facts
- Isotype
- IgG2a
- Host species
- Mouse
- Storage buffer
pH: 7.2 - 7.4
Constituents: 100% PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| WB | IHC-P | ICC/IF | |
|---|---|---|---|
| Human | Tested | Not recommended | Not recommended |
| Mouse | Tested | Not recommended | Not recommended |
| Rat | Tested | Not recommended | Not recommended |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
IHC-P
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
ICC/IF
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
Target data
Function
Belongs to an adhesion system, probably together with the E-cadherin-catenin system, which plays a role in the organization of homotypic, interneuronal and heterotypic cell-cell adherens junctions (AJs) (By similarity). Nectin- and actin-filament-binding protein that connects nectin to the actin cytoskeleton (PubMed:11024295). May play a key role in the organization of epithelial structures of the embryonic ectoderm (By similarity). Essential for the organization of adherens junctions (PubMed:30463011).
Additional Targets
AFDN, AFDN, AFDN, AFDN, AFDN
Alternative names
AF6, MLLT4, AFDN, Afadin, ALL1-fused gene from chromosome 6 protein, Afadin adherens junction formation factor, Protein AF-6
Recommended products
Mouse Monoclonal I Afadin antibody. Carrier free. Suitable for WB and reacts with Human, Mouse, Rat samples.
Key facts
- Isotype
- IgG2a
- Host species
- Mouse
- Storage buffer
pH: 7.2 - 7.4
Constituents: 100% PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- 35/AF6 H1L1
- Purification technique
- Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- Do Not Freeze
Notes
ab300636 is a carrier free version of Anti-AF6 antibody [35/AF6] ab300635.
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 batch production
For more information, read more on recombinant antibodies.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
AF6 also known as Afadin or anti-i is a protein involved in cell signaling and adhesion mechanisms. It weighs approximately 180 kDa and plays a role in the formation of tight junctions influencing cellular architecture and communication. AF6 is expressed in a variety of tissues including epithelial cells indicating its involvement in maintaining tissue integrity. The protein localizes at cellular junctions where it interacts with other junctional components facilitating connections between adjacent cells.
- Biological function summary
AF6 contributes to cell-to-cell contact by organizing cytoskeletal elements and signaling elements. It exists within the nectin-afadin complex which is critical for the regulation of tight junctions and adherens junctions. This positioning allows AF6 to modulate processes like cell migration and tissue morphogenesis. Through its interactions with actin filaments and other junctional proteins AF6 has a significant role in maintaining proper cellular organization and stability.
- Pathways
AF6 participates actively in the Ras signaling and Rap1 pathways. Within the Ras pathway AF6 associates with proteins like Ras and Rap1 which regulate various cellular outcomes including proliferation and differentiation. AF6's involvement with these pathways enhances its role in influencing cell dynamics and signaling decisions. AF6's function in these signaling pathways underlines its participation in normal cell function and development.
- Associated diseases and disorders
AF6 has links to cancer progression and neurological disorders such as autism. The protein’s interaction with Ras influences oncogenic signaling which can lead to aberrant cell growth and metastasis in cancers. Additionally disruptions in AF6 function or expression levels have associations with autism spectrum disorders as AF6 affects neural cell adhesion and signaling pathways. The protein’s interactions with junctional proteins like cadherins and nectins in diseased states suggest its potential as a therapeutic target.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
2 product images
![Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (ab300636), expandable thumbnail](https://content.abcam.com/products/images/af6-antibody-35-af6-bsa-and-azide-free-ab300636--western-blot-img404511.jpg "Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (ab300636)")
Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (ab300636)
This data was developed using Anti-AF6 antibody [35/AF6] ab300635, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Lysates were freshly made and used immediately to minimize protein degradation.
The identity of the lower MW band at approximately 20kDa is unknown.
Exposure time: 3 minutesAll lanes: Western blot - Anti-AF6 antibody [35/AF6] (Anti-AF6 antibody [35/AF6] ab300635) at 1/1000 dilution
Lane 1: 293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg
Lane 3: PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg
SecondaryAll lanes: Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution
Observed band size: 207 kDa, 189 kDa, 190 kDa
Exposure time: 3min
![Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (ab300636), expandable thumbnail](https://content.abcam.com/products/images/af6-antibody-35-af6-bsa-and-azide-free-ab300636--western-blot-img404512.jpg "Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (ab300636)")
Western blot - Anti-AF6 antibody [35/AF6] - BSA and Azide Free (ab300636)
This data was developed using Anti-AF6 antibody [35/AF6] ab300635, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression: (PMID: 21478912 and PMID: 24269953)
The lower bands in may be caused by degradation.
Lysates were freshly made and used immediately to minimize protein degradation.
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
All lanes: Western blot - Anti-AF6 antibody [35/AF6] (Anti-AF6 antibody [35/AF6] ab300635) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg
Lane 2: PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), whole cell lysate at 10 µg
Lane 3: MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg
Lane 4: T47D (human ductal breast epithelial tumor epithelial cell), whole cell lysate at 10 µg
SecondaryAll lanes: Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Developed using the ECL technique.
Observed band size: 207 kDa, 190 kDa
Exposure time: 3min
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