Antibodies, Proteins, Kits and Reagents for Life Science

351 792 ₸ Product size

100 µl 351 792 ₸

Order now and get it on Tuesday March 02, 2021

Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19547-12] to ErbB2 / HER2
Suitable for: ICC/IF, IP, Flow Cyt, IHC-P, WB
Reacts with: Mouse, Rat, Human, Recombinant fragment

Product name

Anti-ErbB2 / HER2 antibody [EPR19547-12]
See all ErbB2 / HER2 primary antibodies
Description

Rabbit monoclonal [EPR19547-12] to ErbB2 / HER2
Host species

Rabbit
Specificity

Only reacts with mouse weakly in 4T1 cells

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Mouse Rat Human Recombinant fragment

See all applications and species data

Immunogen

This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human ErbB 2 fragment recombinant protein; human breast cancer lysate; HeLa, A431, SK-BR-3, C6 and PC-12 whole cell lysates. IHC-P: Human breast cancer tissue. ICC/IF: SK-BR-3 cells. Flow Cyt: SK-BR-3 cells. IP: HeLa whole cell 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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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 (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19547-12
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling ErbB 2 with ab214275 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on Human breast cancer is observed [PMID: 18437174].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

All lanes : Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275) at 1/1000 dilution

Lane 1 : 4T1 (Mouse mammary gland carcinoma epithelial cell) whole cell lysate
Lane 2 : 4T1(Mouse mammary gland carcinoma epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 137 kDa
Observed band size: 180 kDa
why is the actual band size different from the predicted?

Blocking and diluting buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

Immunofluorescent analysis of 100% methanol-fixed SK-BR-3 (Human mammary gland adenocarcinoma cell line) cells labeling ErbB 2 with ab214275 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing membrane staining on SK-BR-3 cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Western blot - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

All lanes : Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275) at 1/1000 dilution

Lane 1 : Human ErbB 2 fragment recombinant protein
Lane 2 : Human ErbB 4 fragment recombinant protein

Lysates/proteins at 0.02 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 137 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

Human ErbB 2 fragment recombinant protein contains aa955-1255 with a His-Tag®; Human ErbB 4 fragment recombinant protein contains aa1000-1300 with a His-Tag®.
Western blot - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275) at 1/2000 dilution + Human breast cancer lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 137 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight is consist with literature PMID :15741050.
Western blot - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

Lanes 1-2 : Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275) at 1/1000 dilution
Lane 3 : Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275) at 1/5000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : SK-BR-3 (Human mammary gland adenocarcinoma cell line) whole cell lysate

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: 137 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 3 minutes; Lane 2: 3 seconds; Lane 3: 10 seconds.

The molecular weight observed is consistent with the literature PMID :15741050.
Western blot - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

All lanes : Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 137 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight is consist with literature PMID :15741050.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling ErbB 2 with ab214275 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on Human breast cancer is observed [PMID: 18437174].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling ErbB 2 with ab214275 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative control: No staining on normal Human breast [PMID: 15150568].

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

Flow cytometric analysis of 4% paraformaldehyde-fixed SK-BR-3 (Human mammary gland adenocarcinoma cell line) cells labeling ErbB 2 with ab214275 at 1/60 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.
Immunoprecipitation - Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

ErbB 2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab214275 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab214275 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution

Lane 1: HeLa whole cell lysate 10µg (Input).

Lane 2: ab214275 IP in HeLa whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214275 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 seconds.
Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com