重组
RabMAb

Anti-Ret抗体[EPR2871] (ab134100)

概述

  • 产品名称
    Anti-Ret抗体[EPR2871]
    参阅全部 Ret 一抗
  • 描述
    兔单克隆抗体[EPR2871] to Ret
  • 宿主
    Rabbit
  • 经测试应用
    适用于: ELISA, WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human Ret (C terminal). The exact sequence is proprietary.
    Database link: P07949
    (Peptide available as ab219199)

  • 阳性对照
    • WB: Neuro-2a, SH-SY5Y and TT cell lysates and mouse and rat brain tissue lysates. IHC-P: Human thyroid gland carcinoma, human gastric carcinoma and mouse colon tissues. ICC/IF: Neuro-2a cells. FC: Caco-2 cells. IP: Neuro-2a cell lysate.
  • 常规说明

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab134100 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ELISA Use a concentration of 0.5 µg/ml.
WB 1/1000 - 1/10000. Predicted molecular weight: 124 kDa.Can be blocked with Ret peptide (ab219199).
IP 1/10 - 1/100.
IHC-P 1/50 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt 1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/50 - 1/100.

靶标

  • 功能
    Probable receptor with tyrosine-protein kinase activity; important for development.
  • 疾病相关
    Defects in RET may be a cause of colorectal cancer (CRC) [MIM:114500].
    Defects in RET are a cause of Hirschsprung disease (HSCR) [MIM:142623]. HSCR is a genetic disorder of neural crest development characterized by the absence of intramural ganglion cells in the hindgut, often resulting in intestinal obstruction. Occasionally, MEN2A or FMTC occur in association with HSCR.
    Defects in RET are the cause of medullary thyroid carcinoma (MTC) [MIM:155240]. MTC is a rare tumor derived from the C cells of the thyroid. Three hereditary forms are known, that are transmitted in an autosomal dominant fashion: (a) multiple neoplasia type 2A (MEN2A), (b) multiple neoplasia type IIB (MEN2B) and (c) familial MTC (FMTC), which occurs in 25-30% of MTC cases and where MTC is the only clinical manifestation.
    Defects in RET are the cause of multiple neoplasia type 2B (MEN2B) [MIM:162300]. MEN2B is an uncommon inherited cancer syndrome characterized by predisposition to MTC and phaeochromocytoma which is associated with marfanoid habitus, mucosal neuromas, skeletal and ophtalmic abnormalities, and ganglioneuromas of the intestine tract. Then the disease progresses rapidly with the development of metastatic MTC and a pheochromocytome in 50% of cases.
    Defects in RET are a cause of susceptibility to pheochromocytoma (PCC) [MIM:171300]. A catecholamine-producing tumor of chromaffin tissue of the adrenal medulla or sympathetic paraganglia. The cardinal symptom, reflecting the increased secretion of epinephrine and norepinephrine, is hypertension, which may be persistent or intermittent.
    Defects in RET are the cause of multiple neoplasia type 2A (MEN2A) [MIM:171400]; also known as multiple neoplasia type 2 (MEN2). MEN2A is the most frequent form of medullary thyroid cancer (MTC). It is an inherited cancer syndrome characterized by MTC, phaeochromocytoma and/or hyperparathyroidism.
    Defects in RET are a cause of thyroid papillary carcinoma (TPC) [MIM:188550]. TPC is a common tumor of the thyroid that typically arises as an irregular, solid or cystic mass from otherwise normal thyroid tissue. Papillary carcinomas are malignant neoplasm characterized by the formation of numerous, irregular, finger-like projections of fibrous stroma that is covered with a surface layer of neoplastic epithelial cells. Note=Chromosomal aberrations involving RET are found in thyroid papillary carcinomas. Inversion inv(10)(q11.2;q21) generates the RET/CCDC6 (PTC1) oncogene; inversion inv(10)(q11.2;q11.2) generates the RET/NCOA4 (PTC3) oncogene; translocation t(10;14)(q11;q32) with GOLGA5 generates the RET/GOLGA5 (PTC5) oncogene; translocation t(8;10)(p21.3;q11.2) with PCM1 generates the PCM1/RET fusion; translocation t(6;10)(p21.3;q11.2) with RFP generates the Delta RFP/RET oncogene; translocation t(1;10)(p13;q11) with TRIM33 generates the TRIM33/RET (PTC7) oncogene; translocation t(7;10)(q32;q11) with TRIM24/TIF1 generates the TRIM24/RET (PTC6) oncogene. The PTC5 oncogene has been found in 2 cases of PACT in children exposed to radioactive fallout after Chernobyl. A chromosomal aberration involving TRIM27/RFP is found in thyroid papillary carcinomas. Translocation t(6;10)(p21.3;q11.2) with RET. The translocation generates TRIM27/RET and delta TRIM27/RET oncogenes.
    Defects in RET are a cause of renal adysplasia (RADYS) [MIM:191830]; also known as renal agenesis or renal aplasia. Renal agenesis refers to the absence of one (unilateral) or both (bilateral) kidneys at birth. Bilateral renal agenesis belongs to a group of perinatally lethal renal diseases, including severe bilateral renal dysplasia, unilateral renal agenesis with contralateral dysplasia and severe obstructive uropathy.
    Defects in RET are a cause of congenital central hypoventilation syndrome (CCHS) [MIM:209880]; also known as congenital failure of autonomic control or Ondine curse. CCHS is a rare disorder characterized by abnormal control of respiration in the absence of neuromuscular or lung disease, or an identifiable brain stem lesion. A deficiency in autonomic control of respiration results in inadequate or negligible ventilatory and arousal responses to hypercapnia and hypoxemia.
  • 序列相似性
    Belongs to the protein kinase superfamily. Tyr protein kinase family.
    Contains 1 cadherin domain.
    Contains 1 protein kinase domain.
  • 翻译后修饰
    Autophosphorylated on C-terminal tyrosine residues upon ligand stimulation. Dephosphorylated by PTPRJ on Tyr-905, Tyr-1015 and Tyr-1062.
  • 细胞定位
    Membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • C ret antibody
    • Cadherin family member 12 antibody
    • Cadherin related family member 16 antibody
    • CDHF 12 antibody
    • CDHF12 antibody
    • CDHR16 antibody
    • ELKS Fusion gene antibody
    • HSCR 1 antibody
    • HSCR1 antibody
    • Hydroxyaryl protein kinase antibody
    • MEN2A antibody
    • MEN2B antibody
    • MTC 1 antibody
    • MTC1 antibody
    • Multiple endocrine neoplasia and medullary thyroid carcinoma 1 antibody
    • Oncogene RET antibody
    • Proto oncogene tyrosine protein kinase receptor ret antibody
    • Proto-oncogene c-Ret antibody
    • Proto-oncogene tyrosine-protein kinase receptor ret antibody
    • PTC antibody
    • RET antibody
    • RET ELE1 antibody
    • Ret Proto oncogene antibody
    • RET transforming sequence antibody
    • RET_HUMAN antibody
    • RET51 antibody
    • RET9 antibody
    • tyrosine-protein kinase receptor ret antibody
    see all

图片

  • Anti-Ret antibody [EPR2871] (ab134100) at 1/1000 dilution (purified) + TT whole cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 124 kDa
    Observed band size: 155,175 kDa (why is the actual band size different from the predicted?)



    Blocking and dilution buffer: 5% NFDM/TBST
  • Anti-Ret antibody [EPR2871] (ab134100) at 1/5000 dilution (purified) + SH-SY5Y whole cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 124 kDa
    Observed band size: 155,175 kDa (why is the actual band size different from the predicted?)



    Blocking and dilution buffer: 5% NFDM/TBST
  • All lanes : Anti-Ret antibody [EPR2871] (ab134100) at 1/1000 dilution (purified)

    Lane 1 : Mouse brain tissue
    Lane 2 : Rat brain tissue

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 124 kDa
    Observed band size: 155,175 kDa (why is the actual band size different from the predicted?)



    Blocking and dilution buffer: 5% NFDM/TBST
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric carcinoma tissue labelling Ret with purified ab134100 at a dilution of 1/50. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of Neuro-2a cells labelling Ret with purified ab134100 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Flow Cytometry analysis of Caco-2 cells labelling Ret with purified ab134100 at a dilution of 1/120 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • ELISA antigen dose-response curve using purified ab134100 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An Alkaline-Phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
  • All lanes : Anti-Ret antibody [EPR2871] (ab134100) at 1/10000 dilution (unpurified)

    Lane 1 : Neuro-2a cell lysate
    Lane 2 : TT cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 124 kDa

  • All lanes : Anti-Ret antibody [EPR2871] (ab134100) at 1/10000 dilution (unpurified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 124 kDa
    Observed band size: 155 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 minutes


    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling Ret with unpurified ab134100 at a dilution of 1/250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse colon tissue labelling Ret with unpurified ab134100 at a dilution of 1/50 followed by HRP-conjugated goat anti-rabbit IgG (H&L) (ab97051, 1/500). Counter stained with hematoxylin.

  • Overlay histogram showing Caco 2 cells stained with unpurified ab134100 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab134100, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Anti-Ret antibody [EPR2871] (ab134100) at 1/10000 dilution (purified) + Neuro-2a whole cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/2000 dilution

    Predicted band size: 124 kDa
    Observed band size: 155,175 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM /TBST.

    We are unsure about the nature of 40kDa to 60kDa bands, they might be the intracellular fragments of Ret.

  • ab134100 (purified) at 1/40 immunoprecipitating Ret in Neuro-2a whole cell lysate.

    Lane 1 (input): Neuro-2a whole cell lysate (10µg)

    Lane 2 (+): ab134100 + Neuro-2a whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab134100 in Neuro-2a whole cell lysate.

    For western blotting, ab134100 was used at 1/1000 followed by a HRP-conjugated anti-rabbit IgG (specific to the non-reduced form of IgG, 1/1500).

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

    We are unsure about the nature of 40kDa to 60kDa bands, they might be the intracellular fragments of Ret.

文献

This product has been referenced in:
  • Nakaoku T  et al. A secondary RET mutation in the activation loop conferring resistance to vandetanib. Nat Commun 9:625 (2018). Read more (PubMed: 29434222) »
  • Tan L  et al. Expression and copy number gains of the RET gene in 631 early and mid stage non-small cell lung cancer cases. Thorac Cancer 9:445-451 (2018). IHC-P ; Human . Read more (PubMed: 29473341) »

See all 19 Publications for this product

客户评价及客户问答

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (neural cell)
Permeabilization
Yes - 0.5% Triton-X 100
Gating Strategy
Positive based off isotype control
Specification
neural cell
Preparation
Cell harvesting/tissue preparation method: Single cell suspension
Sample buffer: 0.1% BSA 10% FCS
Fixation
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 30 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Colon
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Nov 30 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Brain)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
20 µg
Specification
Brain
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Nov 06 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Neuroblastoma)
Permeabilization
Yes - 0.2% Triton X100 in TBS
Specification
Neuroblastoma
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde
Username

Ms. Simin Li

Verified customer

提交于 Apr 28 2015

Application
ELISA
Sample
Mouse Cell (NB41A3 cells)
Specification
NB41A3 cells
Type
Sandwich (Capture)
Blocking step
3% BSA (Sigma) + 1% FBS in PBST (0.05% Tween-20) as blocking agent for 40 minute(s) · Concentration: 0.1% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Mar 03 2015

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (brain)
Specification
brain
Permeabilization
No
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Dec 31 2014

Application
ELISA
Sample
Mouse Cell (NB41A3 cell line)
Specification
NB41A3 cell line
Type
Sandwich (Detection)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 06 2014

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

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