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ASCL1 抗体 (C-Term)

The 兔多克隆 anti-ASCL1 antibody is suitable to detect ASCL1 in samples from 人, 小鼠和大鼠. It has been validated for WB, ELISA, IHC, ICC 和 IF. The antibody has been independently validated.

Cleavage Under Targets and Release Using Nuclease validation image for anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (C-Term) antibody (ABIN6260073) (ASCL1 抗体 (C-Term))

CUT&RUN

Western blot analysis of ASCL1 using RAW264. (ASCL1 抗体 (C-Term))

ABIN6278423 at 1/100 staining Human gastric tissue by IHC-P. (ASCL1 抗体 (C-Term))

IHC

3 images

(1 validation)

产品编号 ABIN6260073

发货至: 中国

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Quick Overview for ASCL1 抗体 (C-Term) (ABIN6260073)

抗原

See all ASCL1 抗体

ASCL1 (Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1))

适用

人, 小鼠, 大鼠

宿主

兔

克隆类型

多克隆

标记

This ASCL1 antibody is un-conjugated

应用范围

Western Blotting (WB), ELISA, Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF)

抗原表位
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C-Term

特异性

ASCL1 Antibody detects endogenous levels of total ASCL1.

预测反应

Pig,Bovine,Sheep,Rabbit,Dog,Chicken,Xenopus

纯化方法

The antiserum was purified by peptide affinity chromatography using SulfoLink^TM Coupling Resin (Thermo Fisher Scientific).

免疫原

A synthesized peptide derived from human ASCL1, corresponding to a region within C-terminal amino acids.

亚型

IgG
anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (AA 151-236) antibody
ASCL1 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC (p), IF (cc), IF (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated

anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (AA 137-236) antibody
ASCL1 适用: 人 WB, ELISA 宿主: 小鼠 Monoclonal 3D3 unconjugated

anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) antibody
ASCL1 适用: 人 ELISA, IHC 宿主: 兔 Polyclonal unconjugated

anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (N-Term) antibody
ASCL1 适用: 人, 小鼠, 大鼠, Cow, 豚鼠, 犬, 兔 WB, IHC 宿主: 兔 Polyclonal unconjugated

anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (AA 84-236) antibody
ASCL1 适用: 人 WB, IHC, ICC, IP 宿主: 兔 Polyclonal unconjugated

anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (AA 137-236) antibody
ASCL1 适用: 人 WB, ELISA, IF 宿主: 小鼠 Monoclonal 2D9 unconjugated

anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (AA 1-236) antibody
ASCL1 适用: 人 WB, ELISA, IHC 宿主: 兔 Polyclonal unconjugated

anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (AA 48-97) antibody
ASCL1 适用: 人, Cow, 豚鼠, 猴 WB, IHC, IHC (p) 宿主: 兔 Polyclonal unconjugated

anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (AA 137-236) antibody
ASCL1 适用: 人 WB, ELISA 宿主: 小鼠 Polyclonal unconjugated

anti-Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1) (AA 79-91) antibody
Verified ASCL1 适用: 人, 小鼠, 大鼠 WB, ELISA, IF 宿主: 山羊 Polyclonal unconjugated

应用备注

WB 1:1000-3000, IHC 1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000

限制

仅限研究用
生效 #104600 (Cleavage Under Targets and Release Using Nuclease)

by

Cantù Lab, Gene Regulation during Development and Disease, Linköping University

No.

#104600

日期

2026.03.01

抗原

ASCL1

Lot Number

35Y1322

Method validated

Cleavage Under Targets and Release Using Nuclease

Positive Control

Anti H3K4me3 (ABIN6971977)

Negative Control

Guinea Pig anti-Rabbit IgG (Heavy & Light Chain) Antibody - Preadsorbed (ABIN101961)

Notes

Passed. ABIN6260073 allows for ASCL1 targeted digestion using CUT&RUN in human neuroendocrine cancer cells.

Validation Images

1+2. Two replicated alignment tracks from CUT&RUN targeting ASCL1 in neuroendocrine cancer organoids using anti-ASCL1 antibody ABIN6260073. 3. Positive Control: Alignment tracks from CUT&RUN targeting H3K3me3 in neuroendocrine cancer organoids using anti-H3K4me3 antibody ABIN6971977. 4. Negative Control: Alignment tracks from CUT&RUN targeting and unspecific IgG.
Full Methods
Primary Antibody

ABIN6260073

Secondary Antibody

Full Protocol

Organoid dissociation into single cells and nuclear extraction. Harvest 200,000 neuroendocrine cancer (NEC) cells per sample. Collect cells in PBS with 0.1% BSA.
Centrifuge cell solution for 5 min at 400 × g at RT. Remove the liquid carefully.
Gently resuspend cells in 1 mL of Nuclear Extraction Buffer (20 mM HEPES-KOH pH 8.2, 20% glycerol, 0.05% IGEPAL, 0.5 mM spermidine, 10 mM KCl, 1× PMSF).
Pellet the nuclei at 800 × g for 5 min. Repeat the NE washes for a total of three washes.
Resuspend the nuclei in 20 µL NE Buffer per sample.
Concanavalin A beads preparation
Prepare 20 µL magnetic Concanavalin A Beads (antibodies-online, ABIN6923139) by 3 washes with 1 mL Binding Buffer (20 mM HEPES pH 7.5, 10 mM KCl, 1 mM CaCl2, 1 mM MnCl2). Gently resuspend the ConA Beads in a volume of Binding Buffer corresponding to the original volume of bead slurry, i.e. 20 µL per sample.
Nuclei immobilization – binding to Concanavalin A beads
Carefully resuspend the nuclei suspension and add 20 µL of the Con A beads in Binding Buffer to the cell suspension for each sample. Let nuclei bind to beads for 15 min at 4 °C with rotation.
Remove the supernatant and resuspend the beads in 200 µL Wash Buffer per sample and divide the cell suspension into separate 200 µL PCR tubes, one for each sample (replicate or antibody).
Remove the supernatant and resuspend the beads in 200 µL of EDTA wash buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 0.5 mM spermidine, 1× PMSF, 2 mM EDTA).
Incubate 5 min at RT.
Primary antibody binding
Prepare 200 µL antibody solution (Wash Buffer with 0.025% digitonin, 2 mM EDTA, and 0.1% BSA) per sample.
Add 2 µL antibody to the respective tube, corresponding to a 1:100 dilution.
Incubate at 4 °C overnight.
Wash with 200 µL of Wash Buffer containing 0.025% digitonin for a total of five washes.
pAG-MNase binding
Prepare 200 µL/sample Wash Buffer containing 0.025% digitonin and 120 ng pAG-MNase/sample. Resuspend beads in pAG-MNase premix.
Incubate 45 min at 4 °C.
Wash with 200 µL of Wash Buffer containing 0.025% digitonin for a total of five washes.
MNase digestion and release of pAG-MNase-antibody-chromatin complexes
Place PCR tubes on ice and allow to chill.
Resuspend in 50 µL of prechilled 2 mM CaCl2 in Wash Buffer mix and incubate on ice for exactly 30 min.
Add 3 µL EDTA (250 mM)-EGTA (250 mM) STOP-Mix per sample, and transfer prerelease into new tubes.
Resuspend the remaining beads in 50 µL of 1× Urea Buffer (8.5 M urea, 100 mM NaCl, 2 mM EGTA, 2 mM EDTA, 0.5% IGEPAL).
Incubate the samples for 45 min at RT.
Remove the urea release from the beads and combine it with the prerelease.
Inactivate the release by adding 96 µL 10 mM Tris, 2 µL SDS 10%, and 2 µL Proteinase K per sample, then incubate at 50 °C for 1 h.
DNA clean-up and sequencing
Clean up DNA by standard phenol-chloroform extraction with overnight precipitation.
Prepare libraries using KAPA HyperPrep Kit with KAPA Dual-Indexed adapters according to protocol.
Sequence samples on an Illumina NextSeq 500 sequencer, using a NextSeq 500/550 High Output Kit v2.5 (75 cycles), 36 bp PE.
Alignment
Trim reads using bbTools bbduk (BBMap - Bushnell B. - sourceforge.net/projects/bbmap/) to remove adapters, artifacts, and repeat sequences.
Aligned reads were mapped to the hg38 human genome using bowtie with options -m 1 -v 0 -I 0 -X 500.
Use SAMtools to convert SAM files to BAM files and remove duplicates.
Use BEDtools genomecov to produce BedGraph files.

Experimental Notes
状态

Liquid

浓度

1 mg/mL

缓冲液

Rabbit IgG in phosphate buffered saline , pH 7.4, 150 mM NaCl, 0.02 % sodium azide and 50 % glycerol.

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

-20 °C

储存方法

Store at -20 °C. Stable for 12 months from date of receipt.

有效期

12 months
抗原

ASCL1 (Achaete-Scute Complex Homolog 1 (Drosophila) (ASCL1))

别名

ASCL1

背景

Description: Transcription factor that plays a key role in neuronal differentiation: acts as a pioneer transcription factor, accessing closed chromatin to allow other factors to bind and activate neural pathways. Directly binds the E box motif (5'-CANNTG-3') on promoters and promotes transcription of neuronal genes. The combination of three transcription factors, ASCL1, POU3F2/BRN2 and MYT1L, is sufficient to reprogram fibroblasts and other somatic cells into induced neuronal (iN) cells in vitro. Plays a role at early stages of development of specific neural lineages in most regions of the CNS, and of several lineages in the PNS. Essential for the generation of olfactory and autonomic neurons. Acts synergistically with FOXN4 to specify the identity of V2b neurons rather than V2a from bipotential p2 progenitors during spinal cord neurogenesis, probably through DLL4-NOTCH signaling activation.
Gene: ASCL1

分子量

25 kDa

基因ID

429

UniProt

P50553

途径

Dopaminergic Neurogenesis