Recombinant Anti-ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284) antibody [EPR19547] - BSA and Azide (ab251328)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19547] to ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284) - BSA and Azide free
- Suitable for: Flow Cyt (Intra), Dot blot, WB, IP, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284) antibody [EPR19547] - BSA and Azide
See all ErbB 2+ErbB 4 primary antibodies -
Description
Rabbit monoclonal [EPR19547] to ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284) - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), Dot blot, WB, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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General notes
ab251328 is the carrier-free version of ab201013.
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19547 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab251328 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Dot blot |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 180 kDa (predicted molecular weight: 138, 147 kDa).
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
Dot blot
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 180 kDa (predicted molecular weight: 138, 147 kDa). |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Target
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Cellular localization
ErbB 2: Cytoplasm. Nucleus and Cell membrane. Cytoplasm, perinuclear region. Nucleus. Translocation to the nucleus requires endocytosis, probably endosomal sorting and is mediated by importin beta-1/KPNB1. ErbB 4: Membrane and Nucleus. Following proteolytical processing E4ICD (E4ICD1 or E4ICD2 generated from the respective isoforms) is translocated to the nucleus. Significantly more E4ICD2 than E4ICD1 is found in the nucleus. E4ICD2 colocalizes with YAP1 in the nucleus. -
Database links
- Entrez Gene: 2064 Human
- Entrez Gene: 2066 Human
- Omim: 164870 Human
- Omim: 600543 Human
- SwissProt: P04626 Human
- SwissProt: Q15303 Human
- Unigene: 390729 Human
- Unigene: 446352 Human
Images
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All lanes : Anti-ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284) antibody [EPR19547] (ab201013) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes
Lane 3 : Untreated A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 4 : A431 (Human epidermoid carcinoma cell line) whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 138, 147 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?This data was developed using ab201013, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-2: 5 seconds; Lanes 3-4: 10 seconds.
This target could be induced to increase by EGF as proved by literatures (PMID: 18945363; 17030621).
The MW is also consistent with some literature (PMID: 18180459).
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All lanes : Anti-ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284) antibody [EPR19547] (ab201013) at 1/5000 dilution
Lane 1 : Untreated SK-BR-3 (human mammary gland adenocarcinoma ) whole cell lysate
Lane 2 : SK-BR-3(human mammary gland adenocarcinoma ) whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 138, 147 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsThis data was developed using ab201013, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
This target could be induced by EGF as shown by literature (PMID: 18945363; 17030621).
The MW is also consistant with that shown within literature (PMID: 18180459).
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This data was developed using ab201013, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ERBB2 / HER2 (phospho Y1248) + ERBB4 / HER4 (phospho Y1284) with ab201013 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing increased cytoplasmic staining after EGF treatment (200ng/ml; 30min). The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red). The negative controls are as follows:- -ve control 1: ab201013 at 1/100 dilution followed by ab150120 at 1/1000 dilution. -ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
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This data was developed using ab201013, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ERBB2 / HER2 (phospho Y1248) + ERBB4 (phospho Y1284) with ab201013 at 1/50 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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This data was developed using ab201013, the same antibody clone in a different buffer formulation.ERBB2 / HER2 (phospho Y1248) + ERBB4 / HER4 (phospho Y1284) were immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes, with ab201013 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201013 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 starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes 10µg (Input). Lane 2: ab201013 IP in HeLa whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes. Lane 3: Rabbit IgG,monoclonal [EPR25A]- Isotype Control (ab172730) instead of ab201013 in HeLa whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 1 second.
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This data was developed using ab201013, the same antibody clone in a different buffer formulation.ERBB2 / HER2 (phospho Y1248) + ERBB4 / HER4 (phospho Y1284) were immunoprecipitated from 0.35 mg of A431 (Human epidermoid carcinoma cell line) whole cell lysate, starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes, with ab201013 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201013 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution. Lane 1: A431 whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes 10µg (Input). Lane 2: ab201013 IP in A431 whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab201013 in A431 whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 1 second.
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
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Datasheet download
Certificate of Compliance
References (0)
ab251328 has not yet been referenced specifically in any publications.