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Rabbit Recombinant Multiclonal Nectin-4 antibody. Suitable for Flow Cyt, WB, IHC-P, IP and reacts with Human samples.

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Images

Flow Cytometry - Anti-Nectin-4 antibody [RM1257] (AB322927), expandable thumbnail
  • Flow Cytometry - Anti-Nectin-4 antibody [RM1257] (AB322927), expandable thumbnail
  • Immunoprecipitation - Anti-Nectin-4 antibody [RM1257] (AB322927), expandable thumbnail
  • Immunoprecipitation - Anti-Nectin-4 antibody [RM1257] (AB322927), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin-4 antibody [RM1257] (AB322927), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Multiclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow CytWBIHC-PIPICC/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

Tested
Tested

Species

Human

Dilution info

1/50

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

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.

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Target data

Function

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.

Alternative names

Rabbit Recombinant Multiclonal Nectin-4 antibody. Suitable for Flow Cyt, WB, IHC-P, IP and reacts with Human samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Multiclonal

Immunogens
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

RM1257

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

This product is a recombinant multiclonal 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.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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.

10 product images

  • Flow Cytometry - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Flow Cytometry - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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 Cytometry - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Flow Cytometry - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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).

  • Immunoprecipitation - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Immunoprecipitation - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Exposure time: 15s

  • Immunoprecipitation - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Immunoprecipitation - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Exposure time: 50s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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

  • Western blot - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Western blot - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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

    Secondary

    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

  • Western blot - Anti-Nectin-4 antibody [RM1257] (ab322927), expandable thumbnail

    Western blot - Anti-Nectin-4 antibody [RM1257] (ab322927)

    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

    Secondary

    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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Product protocols

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