MA5-13040

antibody from Invitrogen Antibodies

Targeting: ERBB3 HER3, LCCS2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-13040

Provider

Invitrogen Antibodies

Product name

ErbB3 Monoclonal Antibody (2B5)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

MA5-13040 targets HER-3 in IP and WB applications and shows reactivity with Human, mouse, and Rat samples.

Antibody clone number

2B5

Concentration

0.2 mg/mL

References

Submitted references

Interaction with epsin 1 regulates the constitutive clathrin-dependent internalization of ErbB3.

Szymanska M, Fosdahl AM, Raiborg C, Dietrich M, Liestøl K, Stang E, Bertelsen V
Biochimica et biophysica acta 2016 Jun;1863(6 Pt A):1179-88

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Soluble-E-cadherin activates HER and IAP family members in HER2+ and TNBC human breast cancers.

Brouxhon SM, Kyrkanides S, Teng X, O'Banion MK, Clarke R, Byers S, Ma L
Molecular carcinogenesis 2014 Nov;53(11):893-906

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Oligomerization of the Nrdp1 E3 ubiquitin ligase is necessary for efficient autoubiquitination but not ErbB3 ubiquitination.

Printsev I, Yen L, Sweeney C, Carraway KL 3rd
The Journal of biological chemistry 2014 Mar 21;289(12):8570-8

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HER3, p95HER2, and HER2 protein expression levels define multiple subtypes of HER2-positive metastatic breast cancer.

Lipton A, Goodman L, Leitzel K, Cook J, Sperinde J, Haddad M, Köstler WJ, Huang W, Weidler JM, Ali S, Newton A, Fuchs EM, Paquet A, Singer CF, Horvat R, Jin X, Banerjee J, Mukherjee A, Tan Y, Shi Y, Chenna A, Larson J, Lie Y, Sherwood T, Petropoulos CJ, Williams S, Winslow J, Parry G, Bates M
Breast cancer research and treatment 2013 Aug;141(1):43-53

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Phosphoinositide 3-kinase pathway activation in phosphate and tensin homolog (PTEN)-deficient prostate cancer cells is independent of receptor tyrosine kinases and mediated by the p110beta and p110delta catalytic subunits.

Jiang X, Chen S, Asara JM, Balk SP
The Journal of biological chemistry 2010 May 14;285(20):14980-14989

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The constitutive activity of epidermal growth factor receptor vIII leads to activation and differential trafficking of wild-type epidermal growth factor receptor and erbB2.

Zeineldin R, Ning Y, Hudson LG
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society 2010 Jun;58(6):529-41

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The ectodomain shedding of E-cadherin by ADAM15 supports ErbB receptor activation.

Najy AJ, Day KC, Day ML
The Journal of biological chemistry 2008 Jun 27;283(26):18393-401

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Activation of nonreceptor tyrosine kinase Bmx/Etk mediated by phosphoinositide 3-kinase, epidermal growth factor receptor, and ErbB3 in prostate cancer cells.

Jiang X, Borgesi RA, McKnight NC, Kaur R, Carpenter CL, Balk SP
The Journal of biological chemistry 2007 Nov 9;282(45):32689-98

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The ErbB4 receptor in fetal rat lung fibroblasts and epithelial type II cells.

Liu W, Zscheppang K, Murray S, Nielsen HC, Dammann CE
Biochimica et biophysica acta 2007 Jul;1772(7):737-47

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ErbB4 regulates fetal surfactant phospholipid synthesis in primary fetal rat type II cells.

Zscheppang K, Liu W, Volpe MV, Nielsen HC, Dammann CE
American journal of physiology. Lung cellular and molecular physiology 2007 Aug;293(2):L429-35

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ErbB receptor dimerization, localization, and co-localization in mouse lung type II epithelial cells.

Zscheppang K, Korenbaum E, Bueter W, Ramadurai SM, Nielsen HC, Dammann CE
Pediatric pulmonology 2006 Dec;41(12):1205-12

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Adult rat otic placode-derived neurons and sensory epithelium express all four erbB receptors: a role in regulating vestibular ganglion neuron viability.

Matsunaga T, Davis JG, Greene MI
DNA and cell biology 2001 Jun;20(6):307-19

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of HER-3 using HER-3 Monoclonal Antibody (Product # MA5-13040) on T47D Cells and MAD109 Cells.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockout of ErbB3 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR764358_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of ErbB3 was performed by loading 60 µg of T-47D wild type (Lane 1), T-47D Cas9 (Lane 2) andT-47D ERBB3 KO (Lane 3) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ 3 to 8%, Tris-Acetate, 1.0 mm, Mini Protein Gel (Product # EC6695BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with ErbB3 Monoclonal Antibody (2B5) (Product # MA5-13040, 1:100 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10.000 dilution) using the iBright™ FL 1500 (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to ErbB3. Uncharacterized bands was observed in wild type and Cas9 lanes at ~71 kDa and ~150 kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of HER-3 was done on 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HER-3 (2B5) Mouse Monoclonal Antibody (Product # MA5-13040) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300).Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of HER-3 using HER-3 Monoclonal Antibody (Product # MA5-13040) on denatured Human T47D Cells.