重组Anti-HMGA1抗体[EPR22421-101] - ChIP Grade (ab252930)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22421-101] to HMGA1 - ChIP Grade
- Suitable for: ChIP, Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-HMGA1抗体[EPR22421-101] - ChIP Grade
参阅全部 HMGA1 一抗 -
描述
兔单克隆抗体[EPR22421-101] to HMGA1 - ChIP Grade -
宿主
Rabbit -
特异性
The Mouse recommendation is based on IP and ChIP results. We do not guarantee WB and IHC for Mouse.
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经测试应用
适用于: ChIP, Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
不适用于: IHC-Fr -
种属反应性
与反应: Mouse, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HT-1080, NCCIT, HCT116 whole cell lysate. Human testis tissue lysate and Human colon cancer tissue lysate, MDA-MB-231 nuclear and membrane fraction lysates, MDA-MB-231 and HCT 116 whole cell lysates prepared in 1% SDS Hot lysis method, MDA-MB-231 and HCT 116 whole cell lysates prepared in RIPA lysis method. IP: HCT116 whole cell lysate, EL4 (mouse). IHC-P: Human testis tissue, Human pancreas cancer and lung cancer tissue. ICC/IF: NCCIT cells. Flow Cyt (intra): NCCIT cells. ChIP: EL4 cells.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR22421-101 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab252930于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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ChIP |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
1/600.
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WB |
1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 12 kDa).
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IHC-P |
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/500.
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IP |
1/30.
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说明 |
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ChIP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
1/600. |
WB
1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 12 kDa). |
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
IP
1/30. |
靶标
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功能
HMG-I/Y bind preferentially to the minor groove of A+T rich regions in double stranded DNA. It is suggested that these proteins could function in nucleosome phasing and in the 3'-end processing of mRNA transcripts. They are also involved in the transcription regulation of genes containing, or in close proximity to A+T-rich regions. -
疾病相关
Note=A chromosomal aberration involving HMGA1 is found in pulmonary chondroid hamartoma. Translocation t(6;14)(p21;q23-24) with RAD51L1. -
序列相似性
Belongs to the HMGA family.
Contains 3 A.T hook DNA-binding domains. -
翻译后修饰
Constitutively phosphorylated on two or three sites. Phosphorylated upon DNA damage, probably by ATM or ATR. Hyperphosphorylated at early stages of apoptosis, followed by dephosphorylation and methylation, which coincides with chromatin condensation. Isoform HMG-Y can be phosphorylated by HIPK2.
HMG-Y is not methylated.
Methylation at Arg-58 is mutually exclusive with methylation at Arg-60. -
细胞定位
Nucleus. Chromosome. - Information by UniProt
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数据库链接
- Entrez Gene: 3159 Human
- Entrez Gene: 15361 Mouse
- Omim: 600701 Human
- SwissProt: P17096 Human
- SwissProt: P17095 Mouse
- Unigene: 518805 Human
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别名
- High mobility group (nonhistone chromosomal) protein isoforms I and Y antibody
- High mobility group AT hook 1 antibody
- High mobility group AT-hook protein 1 antibody
see all
图片
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Chromatin was prepared from EL4 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab252930 (red) or 5 μg of rabbit normal IgG (gray) was added to 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
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All lanes : Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (ab252930) at 1/1000 dilution
Lane 1 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method
Lane 2 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method
Lane 3 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method
Lane 4 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 12 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 40 secondsBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
We recommend using 1% SDS Hot lysis prepare method to get desired Western Blot results.
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Immunohistochemical analysis of paraffin-embedded human testis tissue labeling HMGA1 with ab252930 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in germ cells of human testis (PMID:17935122) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells labeling HMGA1 with ab252930 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in NCCIT cell line. Low expression cell line: MCF7(PMID: 12640109). The nuclear counter stain is DAPI (blue). Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.
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HMGA1 was immunoprecipitated from 0.35 mg HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate with ab252930 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab252930 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.
Lane 1: HCT 116 whole cell lysate 10 µg (Input).
Lane 2: ab252930 IP in HCT 116 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252930 in HCT 116 whole cell lysate.Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 40 seconds. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell, Left) / NCCIT (Human pluripotent embryonic carcinoma epithelial cell, Right) cell lines labeling HMGA1 with ab252930 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. Weak expression control: MCF7 (PMID:12640109).
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All lanes : Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (ab252930) at 1/1000 dilution
Lane 1 : HT-1080 (human fibrosarcoma epithelial cell), whole cell lysate
Lane 2 : NCCIT (human pluripotent embryonic carcinoma epithelial cell), whole cell lysate
Lane 3 : HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate
Lane 4 : Human colon cancer tissue lysate
Lane 5 : Human testis tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 12 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 136 secondsBlocking buffer and concentration 5% NFDM/TBST
Diluting buffer and concentration 5% NFDM/TBSTThe 17-kDa band observed is consistent with what has been described in the literature
(PMID:27045138).Low expression of HMGA1 is expected in the testis (lane 5).
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Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue labeling HMGA1 with ab252930 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human pancreas cancer (PMID: 18473350) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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All lanes : Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (ab252930) at 1/1000 dilution
Lane 1 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) nuclear fraction lysate
Lane 2 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) cytoplasm fraction lysate
Lane 3 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) membrane fraction lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 12 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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HMGA1 was immunoprecipitated from 0.35mg of EL4 (mouse lymphoma T lymphocyte) whole cell lysate with ab252930 at 1/30 dilution.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes : Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (ab252930) at 1/1000 dilution
Lane 1 : EL4 (mouse lymphoma T lymphocyte) whole cell lysate
Lane 2 : ab252930 IP in EL4 (mouse lymphoma T lymphocyte) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab252930 in EL4 (human epidermoid carcinoma) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 180 seconds -
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling HMGA1 with ab252930 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human lung cancer (PMID:25344216) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (4)
ab252930 被引用在 4 文献中.
- Liang G et al. lncRNA IGF2-AS Regulates Nucleotide Metabolism by Mediating HMGA1 to Promote Pyroptosis of Endothelial Progenitor Cells in Sepsis Patients. Oxid Med Cell Longev 2022:9369035 (2022). PubMed: 35082972
- Rajamanickam S et al. Targeting aberrant replication and DNA repair events for treating breast cancers. Commun Biol 5:493 (2022). PubMed: 35610507
- Nakayama J et al. Identification of two cancer stem cell-like populations in triple-negative breast cancer xenografts. Dis Model Mech 15:N/A (2022). PubMed: 35611554
- Pádua D et al. HMGA1 Has Predictive Value in Response to Chemotherapy in Gastric Cancer. Curr Oncol 29:56-67 (2021). PubMed: 35049679