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NOTCH2 抗体 (N-Term)

Name: NOTCH2 抗体 (N-Term)
Brand: antibodies-online
SKU: ABIN1043688
Availability: OutOfStock
Rating: 5 (1 reviews)

NOTCH2 适用: 人 WB, ELISA 宿主: 兔 Polyclonal unconjugated

Western Blotting validation image for anti-Notch 2 (NOTCH2) (N-Term) antibody (ABIN1043688)

anti-NOTCH 2 antibody was diluted 1:500 to detect NOTCH 2 in human liver tissue.

IHC

2 images

(3 references)

(1 validation)

产品编号 ABIN1043688

发货至: 中国

Datasheet as PDF

抗原 See all NOTCH2 抗体

NOTCH2 (Notch 2 (NOTCH2))
抗原表位
16

15

5

4

3

2

2

2

2

2

2

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1
N-Term
适用
62

55

31

6

5

4

2

1

1

1

1

1
人
宿主
73

5

3

1

1
兔
Compatible Secondaries
克隆类型
70

10

2
多克隆
标记
44

4

4

3

2

2

2

2

2

2

2

2

2

2

2

2

1

1

1

1
This NOTCH2 antibody is un-conjugated
应用范围
48

32

26

26

17

16

14

12

7

6

4

2

2

1

1
Western Blotting (WB), ELISA

特异性

This antiserum is directed against human NOTCH 2. Based on sequences, we expect this antibody to react as well with rat and mouse NOTCH 2. Only one amino acid difference (conservative) occurs in mouse and rat. The immunogen epitope is only exposed after gamma secretase cleavage and is not accessible in the uncleaved form.

产品特性

Anti Notch 2 Antibody recognizes Notch 2 that is synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase (S1 cleavage) in the trans-golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved (S2 cleavage) by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called Notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin-dependent gamma-secretase (S3 cleavage) to release the intracellular domain (NICD) from the membrane.

过滤

Sterile filtered

免疫原

This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide corresponding to amino acid residues of human Notch 2 located near the N-terminal sequence of the cleaved N intracellular domain (NICD).
Immunogen Type: Peptide
anti-Notch 2 (NOTCH2) (N-Term) antibody
NOTCH2 适用: 人 WB, ELISA 宿主: 兔 Polyclonal unconjugated

anti-Notch 2 (NOTCH2) (AA 2457-2471) antibody
NOTCH2 适用: 人, 猴 ELISA, IHC, IHC (p) 宿主: 兔 Polyclonal unconjugated

anti-Notch 2 (NOTCH2) antibody
NOTCH2 适用: 人, 小鼠, 大鼠 IHC, IF 宿主: 兔 Polyclonal unconjugated

anti-Notch 2 (NOTCH2) (Asp1733), (cleaved) antibody
NOTCH2 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC 宿主: 兔 Polyclonal unconjugated

anti-Notch 2 (NOTCH2) (C-Term) antibody
NOTCH2 适用: 人, 小鼠 WB, ChIP-seq 宿主: 兔 Polyclonal unconjugated

anti-Notch 2 (NOTCH2) (AA 1501-1700) antibody
NOTCH2 适用: 人, 小鼠 WB, ELISA, IHC (p), IF (cc), IF (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated

anti-Notch 2 (NOTCH2) (Cleaved-Asp1733) antibody
NOTCH2 适用: 人 WB, ELISA, IHC 宿主: 兔 Polyclonal unconjugated

anti-Notch 2 (NOTCH2) (AA 59-250) antibody
NOTCH2 适用: 人 ELISA, IHC, IF 宿主: 兔 Polyclonal unconjugated

anti-Notch 2 (NOTCH2) (AA 2251-2466) antibody
NOTCH2 适用: 人 WB, ELISA, IHC (p), IHC (fro), ICC 宿主: 兔 Polyclonal unconjugated

anti-Notch 2 (NOTCH2) (AA 2242-2471) antibody
NOTCH2 适用: 人 WB, IF 宿主: 兔 Polyclonal unconjugated

应用备注

A strong response was detected by ELISA against the immunizing peptide.This product was assayed against the peptide immunogen in a standard capture ELISA using Peroxidase conjugated anti-Rabbit IgG [H&L] Goat) and ABTS (2,2’-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. A working dilution of 1:30,000 to 1:90,000 is suggested for this product. This product was assayed against the peptide immunogen in a standard capture ELISA using Peroxidase conjugated anti-Rabbit IgG [H&L] (Goat) ( and ABTS (2,2’-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) ( as a substrate for 30 minutes at room temperature. A working dilution of 1:30,000 to 1:90,000 is suggested for this product.

说明

Gene Name: NOTCH2

限制

仅限研究用
生效 #103703 (Western Blotting)

by

AG Pancreatic Development and Stem cell differentiation, Universitätsklinikum Ulm

No.

#103703

日期

2019.06.16

抗原

NOTCH2

Lot Number

Method validated

Western Blotting

Positive Control

pancreatic endoderm cells (day 10)

pancreatic progenitor cells (day 14)

Negative Control

human pluripotent stem cells (day 0)

defnite endoderm cells (day 4)

Notes

ABIN1043688 reveals two protein bands of the expected size and some weaker extraneous bands in cell lysates of human pluripotent stem cells, definite endoderm cells, pancreatic endoderm cells, and pancreatic progenitor cells.

Validation Images

Western blot with ABIN1043688 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). ABIN1043688 was used at a 1:1000 dilution. Vinculin served as loading control.
Full Methods
Primary Antibody

ABIN1043688

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 Tris-HCl pH8.8, 0.1% SDS, 0.1% APS, 0.1% TEMED) for about 120min 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 80min 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-NOTCH2 antibody (antibodies-online, ABIN1043688) diluted 1:1000 respectively in blocking buffer ON at 4°C or
mouse anti-vinculin antibody (SigmaAldrich, V9264) diluted 1:1000 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) with an exposure time of 40min for the ABIN1043688 and 2min for the vinculin loading control.

Experimental Notes

ABIN1043688 revealed some strong unspecific bands. Nevertheless, a band over 250kDa appears to be highly specific, as it is only detected at day 10 (lanes 5 and 6) and day 14 (lanes 7 and 8).
Manual exposure had to be performed due to a defect of the fusion detector in our institute.
ABIN1043688 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.
Interpretation of the results was difficult, as the method of antigen retrieval strongly effects the staining pattern. Antigen retrieval using the citrate buffer at pH6 might be ok, but knockout or knockdown studies are necessary to clearly conclusively show specificity.
状态

Liquid

浓度

70 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
Iervolino, Prosperi, De La Motte, Petrillo, Spagnuolo, DAcierno, Siccardi, Perna, Christensen, Frische, Capasso, Trepiccione: "Potassium depletion induces cellular conversion in the outer medullary collecting duct altering Notch signaling pathway." in: Scientific reports, Vol. 10, Issue 1, pp. 5708, (2020) (PubMed).

Gridley: "Notch signaling and inherited disease syndromes." in: Human molecular genetics, Vol. 12 Spec No 1, pp. R9-13, (2003) (PubMed).

Bertrand, Eckfeldt, Lysholm, LeBien: "Notch-1 and Notch-2 exhibit unique patterns of expression in human B-lineage cells." in: Leukemia, Vol. 14, Issue 12, pp. 2095-102, (2001) (PubMed).
抗原

NOTCH2 (Notch 2 (NOTCH2))

别名

NOTCH 2 (NOTCH2 产品)

别名

AGS2 antibody, HJCYS antibody, hN2 antibody, AI853703 antibody, N2 antibody, notch6 antibody, sb:cb884 antibody, neurogenic locus notch homolog protein 2 antibody, notch 2 antibody, notch 3 antibody, LOC581186 antibody, NOTCH2 antibody, notch3 antibody, Notch2 antibody, notch2 antibody

背景

Anti Notch 2 Antibody recognizes Notch 2 that is synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase (S1 cleavage) in the trans-golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved (S2 cleavage) by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called Notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin-dependent gamma-secretase (S3 cleavage) to release the intracellular domain (NICD) from the membrane.
Synonyms: AGS2 antibody, hN2 antibody, Notch homolog 2 antibody, Notch2 antibody

基因ID

4853, 24041035

UniProt

Q04721

途径

Notch Signaling, Stem Cell Maintenance