Recombinant Anti-Fyn antibody [EPR19636] (ab184276)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19636] to Fyn
- Suitable for: WB, IHC-P, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-Fyn antibody [EPR19636]
See all Fyn primary antibodies -
Description
Rabbit monoclonal [EPR19636] to Fyn -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Human brain lysate; Ramos, HEK-293, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; P0 mouse brain lysate; P0 rat brain lysate; Rat spleen lysate. IHC-P: Human tonsil and cerebral cortex tissues; Mouse spleen tissue; Rat testis tissue. Flow Cyt (intra): RAW 264.7 cells. IP: P0 mouse brain 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
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 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
EPR19636 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab184276 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 |
1/1000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
|
|
IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
IP |
1/40.
|
|
Flow Cyt (Intra) |
1/50.
|
Notes |
---|
WB
1/1000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/40. |
Flow Cyt (Intra)
1/50. |
Target
-
Function
Tyrosine-protein kinase implicated in the control of cell growth. Plays a role in the regulation of intracellular calcium levels, with isoform 2 showing the greater ability to mobilize cytoplasmic calcium in comparison to isoform 1. Required in brain development and mature brain function with important roles in the regulation of axon growth, axon guidance, and neurite extension. Blocks axon outgrowth and attraction induced by NTN1 by phosphorylating its receptor DDC. Phosphorylates RUNX3. -
Tissue specificity
Isoform 1 is highly expressed in the brain. Isoform 2 is expressed in cells of hemopoietic lineages, especially T lymphocytes. -
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
Contains 1 protein kinase domain.
Contains 1 SH2 domain.
Contains 1 SH3 domain. -
Cellular localization
Cell membrane. Present and active in lipid rafts. Present in cell body and along the process of mature and developing oligodendroyctes. - Information by UniProt
-
Database links
- Entrez Gene: 2534 Human
- Entrez Gene: 14360 Mouse
- Entrez Gene: 25150 Rat
- Omim: 137025 Human
- SwissProt: P06241 Human
- SwissProt: P39688 Mouse
- SwissProt: Q62844 Rat
- Unigene: 390567 Human
see all -
Form
This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), YES1 (P07947), SRC (P12931), and LYN (P07948). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request. -
Alternative names
- C syn protooncogene antibody
- Fyn antibody
- FYN oncogene related to SRC FGR YES antibody
see all
Images
-
All lanes : Anti-Fyn antibody [EPR19636] (ab184276) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 cell lysate
Lane 2 : FYN knockout HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Fyn antibody [EPR19636] 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, ab184276 was shown to bind specifically to Fyn. A band was observed at 60 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in FYN knockout cell line ab269630 (knockout cell lysate ab272440). To generate this image, wild-type and FYN knockout HEK-293 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
-
All lanes : Anti-Fyn antibody [EPR19636] (ab184276) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : FYN knockout HEK-293T cell lysate
Lane 3 : Human brain tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 159 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab184276 observed at 60 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab184276 Anti-Fyn antibody [EPR19636] was shown to specifically react with Fyn in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266133 (knockout cell lysate ab257071) was used. Wild-type and Fyn knockout samples were subjected to SDS-PAGE. ab184276 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 10000 dilution for 1 hour at room temperature before imaging.
-
All lanes : Anti-Fyn antibody [EPR19636] (ab184276) at 1/1000 dilution
Lane 1 : Human brain lysate
Lane 2 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
Lane 3 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate
Lane 4 : P0 mouse brain lysate
Lane 5 : P0 rat brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Lanes 2-5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lanes 1 and 2: 3 minutes; Lanes 3,4 and 5: 8 seconds.
-
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on T cells of human tonsil (PMID: 10523617). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human cerebral cortex (PMID: 7544314). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on T cells of mouse spleen (PMID: 7544314, PMID:10523617). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on rat testis (PMID: 7544314). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
All lanes : Anti-Fyn antibody [EPR19636] (ab184276) at 1/1000 dilution
Lane 1 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 3 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
Lane 4 : Rat spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
-
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line labeling Fyn with ab184276 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
-
Fyn was immunoprecipitated from 0.35 mg of P0 mouse brain lysate with ab184276 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab184276 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: P0 mouse brain lysate 10 µg (Input).
Lane 2: ab184276 IP in P0 mouse brain lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184276 in P0 mouse brain lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (5)
ab184276 has been referenced in 5 publications.
- Yang Y et al. FYN regulates cell adhesion at the blood-testis barrier and the apical ectoplasmic specialization via its effect on Arp3 in the mouse testis. Front Immunol 13:915274 (2022). PubMed: 36016954
- Tan C et al. Fyn kinase regulates dopaminergic neuronal apoptosis in animal and cell models of high glucose (HG) treatment. BMC Mol Cell Biol 22:58 (2021). PubMed: 34863087
- Luo X et al. Fyn gene silencing reduces oligodendrocytes apoptosis through inhibiting ERK1/2 phosphorylation in epilepsy. Artif Cells Nanomed Biotechnol 48:298-304 (2020). PubMed: 31852295
- Tang Y et al. Bioinformatic analysis of differentially expressed genes and identification of key genes in EBV-transformed lymphoblasts. Biomed Pharmacother 116:108984 (2019). PubMed: 31129512
- Zambrano S et al. GPRC5b Modulates Inflammatory Response in Glomerular Diseases via NF-?B Pathway. J Am Soc Nephrol 30:1573-1586 (2019). PubMed: 31285284