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JAG1 抗体 (Internal Region)

Name: JAG1 抗体 (Internal Region)
Brand: antibodies-online
SKU: ABIN129524
Availability: OutOfStock
Rating: 5 (1 reviews)

JAG1 适用: 人, 小鼠 WB, ELISA, IHC, FM 宿主: 兔 Polyclonal unconjugated

Western Blotting validation image for anti-Jagged 1 (JAG1) (Internal Region) antibody (ABIN1043767) (JAG1 抗体 (Internal Region))

Western blot using Protein A purified anti-Jagged-1 antibody shows detection of Jagged-1 protein in various whole cell lysates: human brain (lane 1), human kidney (lane 2), human liver (lane 3), and mouse liver (lane 5). (JAG1 抗体 (Internal Region))

Immunohistochemical staining of human cervical cancer tissue (40X magnification) using Protein A purified anti-Jagged-1 antibody. (JAG1 抗体 (Internal Region))

IHC

Immunofluorescence microscopy using Protein A purified anti-Jagged-1 antibody of human corneal epithelial cells. (JAG1 抗体 (Internal Region))

4 images

(2 references)

(1 validation)

产品编号 ABIN129524

发货至: 中国

Datasheet as PDF

抗原 See all JAG1 抗体

JAG1 (Jagged 1 (JAG1))
抗原表位
12

8

7

5

5

3

3

3

2

2

2

2

2

2

2

2

1

1

1

1

1

1

1
Internal Region
适用
85

39

36

1
人, 小鼠
宿主
73

15

3

1
兔
Compatible Secondaries
克隆类型
70

22
多克隆
标记
41

9

8

4

4

4

2

2

2

2

2

2

1

1

1

1

1

1

1

1

1

1
This JAG1 antibody is un-conjugated
应用范围
62

33

25

15

11

11

9

9

6

5

3

2

1

1

1
Western Blotting (WB), ELISA, Immunohistochemistry (IHC), Fluorescence Microscopy (FM)

特异性

This protein A purified antibody is directed against human Jagged-1 protein. The product was purified from mono-specific antiserum by affinity chromatography. A BLAST analysis was used to suggest cross reactivity with Jagged-1 protein from human, chimpanzee, rat and mouse based on 100% homology with the immunizing sequence. Partial reactivity is expected against dog (81%) and Xenopus laevis (85%) based on partial sequence homologies as indicated. Reactivity against homologues from other sources is not known.

产品特性

Anti Jagged 1 Antibody recognizes the jagged 1 protein encoded by the JAG1 gene, that is the human homolog of the Drosophilia jagged protein. Human jagged 1 is the ligand for the receptor notch 1, the latter a human homolog of the Drosophilia jagged receptor notch. Mutations that alter the jagged 1 protein cause Alagille syndrome. Jagged 1 signaling through notch 1 has also been shown to play a role in hematopoies.

纯化方法

purified

过滤

Sterile filtered

免疫原

This protein A purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 110-125 of human Jagged-1protein.
Immunogen Type: Peptide

亚型

IgG
anti-Jagged 1 (JAG1) (Internal Region) antibody
JAG1 适用: 人, 小鼠, 大鼠 WB, ELISA, IF, IHC (p) 宿主: 兔 Polyclonal unconjugated

anti-Jagged 1 (JAG1) (AA 47-212) antibody
JAG1 适用: 人 ELISA, FACS 宿主: 小鼠 Monoclonal 6A9B2 unconjugated

anti-Jagged 1 (JAG1) (AA 836-1047) antibody
JAG1 适用: 人 WB, IHC, IP, ICC 宿主: 兔 Polyclonal unconjugated

anti-Jagged 1 (JAG1) (AA 809-1046) antibody
JAG1 适用: 人 ELISA, IHC, IF 宿主: 兔 Polyclonal unconjugated

anti-Jagged 1 (JAG1) antibody
JAG1 适用: 人 WB, ELISA, IHC, FACS 宿主: 小鼠 Monoclonal J1G53-3 unconjugated

anti-Jagged 1 (JAG1) (AA 470-834) antibody
JAG1 适用: 人 WB, IHC, IP, ICC 宿主: 兔 Polyclonal unconjugated

anti-Jagged 1 (JAG1) (AA 33-250) antibody
JAG1 适用: 人 WB, IHC, IP, ICC 宿主: 兔 Polyclonal unconjugated

anti-Jagged 1 (JAG1) (AA 826-1084) antibody
JAG1 适用: 小鼠 WB, IHC, IP, ICC 宿主: 兔 Polyclonal unconjugated

anti-Jagged 1 (JAG1) (N-Term) antibody
JAG1 适用: 人, 小鼠, 大鼠 WB, ELISA 宿主: 兔 Polyclonal unconjugated

anti-Jagged 1 (JAG1) (C-Term) antibody
Verified JAG1 适用: 人, 小鼠 ELISA, IHC, IF 宿主: 山羊 Polyclonal unconjugated

应用备注

This protein A purified antibody has been tested for use in ELISA, immunohistochemistry, immunofluorescence microscopy and western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 150 kDa in size corresponding to Jagged-1 in mouse liver whole cell lysates, and a 75 kDa band in human brain and kidney lysates.

说明

Gene Name: JAG1

限制

仅限研究用
生效 #103702 (Western Blotting)

by

AG Pancreatic Development and Stem cell differentiation, Universitätsklinikum Ulm

No.

#103702

日期

2019.06.16

抗原

JAG1

Lot Number

28285

Method validated

Western Blotting

Positive Control

human pluripotent stem cells (day 0)

pancreatic progenitor cells (day 14)

Negative Control

definite endoderm cells (day 4)

pancreatic endoderm cells (day 10)

Notes

ABIN1043767 reveals a protein band of the expected MW and some weaker extraneous bands in cell lysates of human pluripotent stem cells and pancreatic progenitor cells.

Validation Images

Western blot with ABIN1043686 on human pluripotent stem cells (1 and 2), definite endoderm cells (3 and 4), pancreatic endoderm cells (5 and 6), and pancreatic progenitors (7 and 8). ABIN1043686 was used at a 1:1000 (30sec exposure) or 1:10000 dilution (5min exposure). Beta-actin served as loading control.
Full Methods
Primary Antibody

ABIN1043767

Secondary Antibody

donkey anti-rabbit HRP-conjugated antibody (GE Healthcare, NA9310V)

Full Protocol

Grow HUES8 in mTeSR1 (STEMCELL Technologies, 85850) at 37°C and 5% O₂, 5% CO₂ in 2ml in a 6-well plate to 90% confluency.
Harvest cells using TrypLE (ThermoFisher Scientific, 12604013) following the manufacturer´s instructions.
Lyse 2x10⁶ cells in 30µl per well cold RIPA buffer (50mM Tris-HCl pH 8.0, 150mM NaCl, 0.1% SDS, 0.5% deoxycholate, 1% TritonX 100 in ddH₂O) supplemented with 1mM PMSF and 1x protease inhibitor (Roche, 11836170001) for 30min on ice with 3x vortexing in 1.5ml microcentrifuge tubes.
Centrifuge tubes at 10000xg for 8min at 4°C.
Transfer supernatant to a new 1.5ml microcentrifuge tube and store at -80°C.
Determine total protein content of the lysates using a Bradford assay (Bio-Rad, 500-0006).
Denature 50µg of total protein for 5min at 95°C in 30µl 1x Laemmli SDS sample buffer and subsequently separate them on a denaturing 7.5% polyacrylamide gel (7.5% Acrylamide, 0.375M Trís-HCl (pH8.8), 0.1% SDS, 0.1% APS, 0.1% TEMED) for about 90min at 120V.
Transfer proteins onto a PVDF membrane (Sigma Aldrich, IPVH00010) with transfer buffer (5.27g Tris, 2.93 g glycerine, 200 ml methanol, fill to 1l ddH₂O) in a semidry western blotting system for 75min at 80mA/gel.
Check transfer of the separated proteins by Ponceau S staining.
Rinse membrane with water.
Wash membrane for 5min with TBST (TBS, 0.02% Tween20).
Block the membrane with 20ml blocking buffer (TBST, 5% milk) on a shaker for 1h at RT.
Rinse membrane 3x with TBST.
Wash membrane on a shaker 3x for 5min with TBST.
Shrink-wrap and incubate membrane with primary
rabbit anti-JAG1 antibody (antibodies-online, ABIN1043767, lot 28285) diluted 1:1000 and 1:10000 respectively in blocking buffer ON at 4°C or
mouse anti-beta actin antibody (Sigma Aldrich, A1978) diluted 1:2000 in blocking buffer at RT for 1h.
Wash membrane 3x for 5min with TBST.
Incubate membrane with secondary
donkey anti-rabbit HRP-conjugated antibody (GE Healthcare, NA9310V) diluted 1:5000 in TBST containing 1% milk for 1h at RT or
donkey anti-mouse HRP-conjugated antibody (GE Healthcare, NA9340V) diluted 1:5000 in TBST containing 1% milk for 1h at RT.
Wash membrane 3x for 5min with TBST.
Reveal protein bands using ECL solution (ThermoScientific, 34076) on a Fusion SL (Vilber) chemiluminescence system. Exposure times for the 1:1000 and 1:10000 dilutions of ABIN1043767 were 30sec and 5min respectively.

Experimental Notes

The bands marked by the arrow in the western blot image using ABIN1043767 at a 1:1000 dilution seem to be specific, as JAG1 showed highest transcriptomic levels at pancreatic progenitor stage (lanes 7 and 8), followed by pluripotent stem cell state (lane 2). Although the immunoblot with ABIN1043767 results in some unspecific bands, the expected band height fits nicely (75kDa). Higher MW bands might potentially represent precursor proteins detectable in all samples but pancreatic progenitor stage samples. In principle specific bands are also detectable with ABIN1043767 at a 1:10000 dilution, although exposition time was very long.
ABIN1043767 was also tested in IHC on 4-5µm FFPE sections of primary human kidney cancer tissue. Epitope retrieval was carried out using Tris-EDTA buffer at pH9.0 (Zytomed, ZUC029-500), EDTA at pH8.0 (Leica, RE7116), or citrate buffer at pH6.1 (Agilent, S169984-2) for 20min in a decloaking chamber. Antigen retrieval at pH8.0 looks ok, assuming a broad expression pattern. However, detection would be rather expected in fewer tubular cells. No staining was observed in glomeruli. Cytoplasmic staining in tubuli is revealed at higher magnification. Note that antigen retrieval influences the staining pattern. All in all, staining needs to be validated for example in protein ablation assays.
状态

Liquid

浓度

1.1 mg/mL

缓冲液

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

储存液

Sodium azide

注意事项

This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

储存条件

4 °C/-20 °C

储存方法

Store vial at 4 °C prior to restoration. For extended storage aliquot contents and freeze at -20 °C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4 °C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

有效期

12 months
Boareto, Jolly, Goldman, Pietilä, Mani, Sengupta, Ben-Jacob, Levine, Onuchic: "Notch-Jagged signalling can give rise to clusters of cells exhibiting a hybrid epithelial/mesenchymal phenotype." in: Journal of the Royal Society, Interface, Vol. 13, Issue 118, (2017) (PubMed).

Yabe, Matsumoto, Tsurumoto, Shindo: "Immunohistological localization of Notch receptors and their ligands Delta and Jagged in synovial tissues of rheumatoid arthritis." in: Journal of orthopaedic science : official journal of the Japanese Orthopaedic Association, Vol. 10, Issue 6, pp. 589-94, (2005) (PubMed).
抗原

JAG1 (Jagged 1 (JAG1))

别名

Jagged 1 (JAG1 产品)

别名

AGS antibody, AHD antibody, AWS antibody, CD339 antibody, HJ1 antibody, JAGL1 antibody, ABE2 antibody, Gsfabe2 antibody, Htu antibody, Ozz antibody, Ser-1 antibody, JAG1 antibody, jag3 antibody, cb243 antibody, jag1 antibody, sb:cb243 antibody, serrateC antibody, jagged1 antibody, serrate antibody, serrate-1 antibody, x-serrate-1 antibody, jagged 1 antibody, jagged 1b antibody, jagged 1a antibody, jagged 1 L homeolog antibody, JAG1 antibody, Jag1 antibody, jag1 antibody, jag1b antibody, jag1a antibody, jag1.L antibody

背景

Anti Jagged 1 Antibody recognizes the jagged 1 protein encoded by the JAG1 gene, that is the human homolog of the Drosophilia jagged protein. Human jagged 1 is the ligand for the receptor notch 1, the latter a human homolog of the Drosophilia jagged receptor notch. Mutations that alter the jagged 1 protein cause Alagille syndrome. Jagged 1 signaling through notch 1 has also been shown to play a role in hematopoies.
Synonyms: Ser 1 antibody, AGS antibody, AHD antibody, AWS antibody, CD 339 antibody, CD339 antibody, CD339 antigen antibody, Headturner antibody, HJ1 antibody, Htu antibody

基因ID

182, 4557679

UniProt

P78504

途径

Notch Signaling, Stem Cell Maintenance