Rabbit Recombinant Multiclonal Nectin-4 antibody. Suitable for Flow Cyt, WB, IHC-P, IP and reacts with Human samples.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Multiclonal
Flow Cyt | WB | IHC-P | IP | ICC/IF | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Not recommended |
Mouse | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Rat | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/50 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
Seems to be involved in cell adhesion through trans-homophilic and -heterophilic interactions, the latter including specifically interactions with NECTIN1. Does not act as receptor for alpha-herpesvirus entry into cells.(Microbial infection) Acts as a receptor for measles virus.
LNIR, PRR4, PVRL4, PVRL4, PRR4, LNIR, NECTIN4, Nectin-4, Ig superfamily receptor LNIR, Nectin cell adhesion molecule 4, Poliovirus receptor-related protein 4
Rabbit Recombinant Multiclonal Nectin-4 antibody. Suitable for Flow Cyt, WB, IHC-P, IP and reacts with Human samples.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Multiclonal
RM1257
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
This product is a recombinant multiclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
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 have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
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.
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.
Flow cytometric analysis of T-47D (human ductal breast epithelial tumor epithelial cell) (Right) / A549 (human lung carcinoma epithelial cell) (Left) cells labelling Nectin-4 with ab322927 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Low expression: A549 (PMID:21901103).
Flow cytometric analysis of DLD-1 (human colorectal adenocarcinoma epithelial cell) (Right) / HeLa (human cervical adenocarcinoma epithelial cell) (Left) cells labelling Nectin-4 with ab322927 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Low expression: HeLa (PMID:21901103).
Nectin-4 was immunoprecipitated from 0.35 mg T-47D whole cell lysate with ab322927 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322927 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate
Lane 2: ab322927 IP in T-47D whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab322927 in T-47D whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-Nectin-4 antibody [RM1257] (ab322927) at 1/30 dilution
All lanes: T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 15s
Nectin-4 was immunoprecipitated from 0.35 mg DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate with ab322927 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322927 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 2: ab322927 IP in DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab322927 in DLD-1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-Nectin-4 antibody [RM1257] (ab322927) at 1/30 dilution
All lanes: DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 50s
Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Nectin-4 with ab322927 at 1/2000 (0.265 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: no staining on human spleen.
The section was incubated with ab322927 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling Nectin-4 with ab322927 at 1/2000 (0.265 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: no staining on human cardiac muscle.
The section was incubated with ab322927 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling Nectin-4 with ab322927 at 1/2000 (0.265 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human skin.
The section was incubated with ab322927 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Nectin-4 with ab322927 at 1/2000 (0.265 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human placenta.
The section was incubated with ab322927 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: Spleen (PMID: 11544254).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Nectin-4 antibody [RM1257] (ab322927) at 1/1000 dilution
Lane 1: Human placenta tissue lysate at 20 µg
Lane 2: Human spleen tissue lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Observed band size: 40-70 kDa, 36 kDa
Exposure time: 180s
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: HeLa and A549 (PMID: 21901103).
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15784625).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Nectin-4 antibody [RM1257] (ab322927) at 1/1000 dilution
Lane 1: HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 3: T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg
Lane 4: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 5: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 6: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 7: A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 40-70 kDa, 36 kDa
Exposure time: 147s
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