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Phospho-HER2 / ErbB2 (Y1248) Recombinant Rabbit Monoclonal Antibody [PSH04-03] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750926

Phospho-HER2 / ErbB2 (Y1248) Recombinant Rabbit Monoclonal Antibody [PSH04-03] - BSA and Azide free

Application

  • WB

  • IF-Cell

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Phospho-HER2 / ErbB2 (Y1248) Recombinant Rabbit Monoclonal Antibody [PSH04-03] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Tyr1248 of Human HER2.

Species Reactivity

Human

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 138 kDa

Positive Control

SK-Br-3 cell lysate, SK-Br-3 starved for 4 hours add 200ng/mL EGF for 15 minutes cell lysate, HeLa starved for 4 hours add 200ng/mL EGF for 15 minutes cell lysate, HeLa cells starved for 4 hours add 200ng/mL EGF for 15 minutes.

Conjugation

unconjugated

Clone Number

PSH04-03

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

HER-2 (also called HER-2/neu, c-erbB2, ERBB2 or neu) is a transmembrane receptor tyrosine kinase. HER-2 is a proto-oncogene, i.e. its activation causes malignant transformation and increases the malignant potential (cell proliferation, invasiveness etc.) of the cells. Amplification of HER-2 gene invariably leads to over-expression of its protein product. The magnitude of over- expression is usually 10-100 folds or even more. Over-expressed HER-2 protein disturbs the HER-receptor family signalling networks, i.e. signalling mediated via EGFR receptor, HER-3 and HER-4. In tumours, HER-2 is over-expressed in 15-25% of primary breast cancers. Metastases usually have the same amplification status as the primary tumours. HER-2 amplification and over-expression are typical features of hormone receptor negative, rapidly growing histologic grade 2-3 tumours. Of the histologic types, Paget’s disease is almost invariably HER-2 positive, whereas only a small minority of lobular and tubular carcinomas shows HER-2 amplification. HER-2 amplification and over-expression can also be found in intestinal type gastric and gastroesophageal carcinomas, ovarian carcinomas, high grade endometrial carcinomas and some salivary duct tumours. Low-level copy number increases have been found also in rare cases of lung tumours. Because of its central importance in breast cancer therapy selection, standardization of HER-2 IHC assays and slide interpretation are of outmost clinical and economical importance.

Background References

1. Fernandez AI et al. Examination of Low ERBB2 Protein Expression in Breast Cancer Tissue. JAMA Oncol. 2022 Apr

2. Augustin JE et al. Targeting the complexity of ERBB2 biology in gastroesophageal carcinoma. Ann Oncol. 2022 Nov

Subcellular Location

Cell membrane, Cell projection, ruffle membrane; Early endosome, Cytoplasm, perinuclear region, Nucleus.

UNIPROT

SwissProt: P04626 Human

Synonyms

Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog antibody

C erb B2/neu protein antibody

CD340 antibody

CD340 antigen antibody

Cerb B2/neu protein antibody

CerbB2 antibody

Erb b2 receptor tyrosine kinase 2 antibody

ErbB-2 proto-oncogene antibody

ERBB2 antibody

ERBB2_HUMAN antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-HER2 / ErbB2 (Y1248) on different lysates with Rabbit anti-Phospho-HER2 / ErbB2 (Y1248) antibody (<a href="/products/HA750926" style="font-weight: bold;text-decoration: underline;">HA750926</a>) at 1/1,000 dilution.<br /><br />Lane 1: SK-Br-3 cell lysate<br />Lane 2: SK-Br-3 starved for 4 hours add 200ng/mL EGF for 15 minutes cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: HeLa starved for 4 hours add 200ng/mL EGF for 15 minutes cell lysate<br />Lane 5: HeLa starved for 4 hours add 200ng/mL EGF for 15 minutes cell lysate, then the membrane treated with λpp for 1 hour<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 138 kDa<br />Observed band size: 180 kDa<br /><br />Exposure time: 24 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750926" style="font-weight: bold;text-decoration: underline;">HA750926</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-HER2 / ErbB2 (Y1248) on different lysates with Rabbit anti-Phospho-HER2 / ErbB2 (Y1248) antibody (HA750926) at 1/1,000 dilution.

    Lane 1: SK-Br-3 cell lysate
    Lane 2: SK-Br-3 starved for 4 hours add 200ng/mL EGF for 15 minutes cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: HeLa starved for 4 hours add 200ng/mL EGF for 15 minutes cell lysate
    Lane 5: HeLa starved for 4 hours add 200ng/mL EGF for 15 minutes cell lysate, then the membrane treated with λpp for 1 hour

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 138 kDa
    Observed band size: 180 kDa

    Exposure time: 24 seconds;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750926) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells starved for 4 hours add 200ng/mL EGF for 15 minutes labeling Phospho-HER2 / ErbB2 (Y1248) with Rabbit anti-Phospho-HER2 / ErbB2 (Y1248) antibody (<a href="/products/HA750926" style="font-weight: bold;text-decoration: underline;">HA750926</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-HER2 / ErbB2 (Y1248) antibody (<a href="/products/HA750926" style="font-weight: bold;text-decoration: underline;">HA750926</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells starved for 4 hours add 200ng/mL EGF for 15 minutes labeling Phospho-HER2 / ErbB2 (Y1248) with Rabbit anti-Phospho-HER2 / ErbB2 (Y1248) antibody (HA750926) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-HER2 / ErbB2 (Y1248) antibody (HA750926) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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