使用敲除细胞株进行验证

Anti-ABCA1抗体[AB.H10] (ab18180)

概述

  • 产品名称
    Anti-ABCA1抗体[AB.H10]
    参阅全部 ABCA1 一抗
  • 描述
    小鼠单克隆抗体[AB.H10] to ABCA1
  • 经测试应用
    适用于: IHC-P, ICC/IF, Flow Cyt, WB, ELISAmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Cow, Human
  • 免疫原

    Recombinant fragment corresponding to Human ABCA1 aa 1800-2260.
    Database link: O95477

  • 阳性对照
    • testis, liver, and brain tissue (negative control: muscle tissue)

应用

Our Abpromise guarantee covers the use of ab18180 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
IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB 1/200 - 1/500. Predicted molecular weight: 254 kDa.

Abcam recommends using BSA as the blocking agent and 80-100 ug of lysate.

ELISA Use at an assay dependent concentration.

Direct ELISA, where a 96-well plate was coated with recombinant ABCA1 C-terminal protein. The antibody was then added to the wells and a goat anti-mouse IgG HRP conjugate was used for colour development. For ABCA1 quantification, a pair of anti-ABCA1 antibodies should be used to formulate a Sandwich ELISA and with a presence of an ABCA1 protein Standard. Optimal dilution to be determined by the end user.

靶标

  • 功能
    cAMP-dependent and sulfonylurea-sensitive anion transporter. Key gatekeeper influencing intracellular cholesterol transport.
  • 组织特异性
    Widely expressed, but most abundant in macrophages.
  • 疾病相关
    Defects in ABCA1 are a cause of high density lipoprotein deficiency type 1 (HDLD1) [MIM:205400]; also known as analphalipoproteinemia or Tangier disease (TGD). HDLD1 is a recessive disorder characterized by absence of high density lipoprotein (HDL) cholesterol from plasma, accumulation of cholesteryl esters, premature coronary artery disease (CAD), hepatosplenomegaly, recurrent peripheral neuropathy and progressive muscle wasting and weakness.
    Defects in ABCA1 are a cause of high density lipoprotein deficiency type 2 (HDLD2) [MIM:604091]; also known as familial hypoalphalipoproteinemia (FHA). HDLD2 is inherited as autosomal dominant trait. It is characterized by moderately low HDL cholesterol, predilection toward premature coronary artery disease (CAD) and a reduction in cellular cholesterol efflux.
  • 序列相似性
    Belongs to the ABC transporter superfamily. ABCA family.
    Contains 2 ABC transporter domains.
  • 结构域
    Multifunctional polypeptide with two homologous halves, each containing an hydrophobic membrane-anchoring domain and an ATP binding cassette (ABC) domain.
  • 翻译后修饰
    Phosphorylation on Ser-2054 regulates phospholipid efflux.
    Palmitoylation by DHHC8 is essential for membrane localization.
  • 细胞定位
    Membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ABC 1 antibody
    • ABC Transporter 1 antibody
    • ABC-1 antibody
    • ABC1 antibody
    • ABCA 1 antibody
    • ABCA1 antibody
    • ABCA1_HUMAN antibody
    • ATP binding Cassette 1 antibody
    • ATP binding cassette sub family A ABC1 member 1 antibody
    • ATP binding cassette sub family A member 1 antibody
    • ATP binding Cassette Transporter 1 antibody
    • ATP-binding cassette 1 antibody
    • ATP-binding cassette sub-family A member 1 antibody
    • ATP-binding cassette transporter 1 antibody
    • CERP antibody
    • Cholesterol efflux regulatory protein antibody
    • FLJ14958 antibody
    • HDLDT1 antibody
    • Membrane bound antibody
    • MGC164864 antibody
    • MGC165011 antibody
    • TD antibody
    • TGD antibody
    see all

图片



  • Predicted band size : 254 kDa
    Western blot analysis of ABCA1 induction by cholesterol (+) in human fibroblast cells from different patients (1,2,3,4). Simultaneous bloting with anti-actin antibody was used for protein loading control. Western blot analysis of ABCA1 induction by cholesterol (+) in human fibroblast cells from different patients (1,2,3,4). Simultaneous blotting with anti-actin antibody was used for protein loading control.
  • Overlay histogram showing HepG2 cells stained with ab18180 (red line). The cells were fixed with 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 (ab18180, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • All lanes : Anti-ABCA1 antibody [AB.H10] (ab18180) at 1/1000 dilution

    Lane 1 : Wild type mouse liver tissue lysate
    Lane 2 : Knockout mouse liver tissue lysate
    Lane 3 : Wild type mouse liver tissue lysate
    Lane 4 : Knockout mouse liver tissue lysate
    Lane 5 : Wild type mouse liver tissue lysate
    Lane 6 : Knockout mouse liver tissue lysate

    Lysates/proteins at 100 µg per lane.


    Predicted band size : 254 kDa
    Observed band size : 254 kDa

    The lower bands are from the secondary anti-mouse antibody reacting to the endogenous mouse antibodies from the tissue.

  • Immunoreactivity of pancreas tissue for ABCA1 protein. Left panel: strong immunopositivity of exocrine glandular cells of the pancreas, especially in the basal region (arrows). Interstitial microvascular cells are apparently negative. Cells within the islet were weakly positive (data not shown). Right panel: the negative control. Hematoxylin counterstain.

文献

This product has been referenced in:
  • Vienken H  et al. Characterization of cholesterol homeostasis in sphingosine-1-phosphate lyase-deficient fibroblasts reveals a Niemann-Pick disease type C-like phenotype with enhanced lysosomal Ca(2+) storage. Sci Rep 7:43575 (2017). WB . Read more (PubMed: 28262793) »
  • Sithu SD  et al. Atherogenesis and metabolic dysregulation in LDL receptor-knockout rats. JCI Insight 2:N/A (2017). WB ; Rat . Read more (PubMed: 28469073) »

See all 58 Publications for this product

客户评价及客户问答

Application
Western blot
Sample
Sheep Cell lysate - whole cell (Ovine Thecal Cells)
Gel Running Conditions
Reduced Denaturing (10% PAG)
Loading amount
10 µg
Specification
Ovine Thecal Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Miss. Barbara Makela

Verified customer

提交于 Jun 22 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Gel Running Conditions
Reduced Denaturing (7.5)
Loading amount
25 µg
Specification
liver
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 24 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary astrocyte culture)
Permeabilization
Yes - 0.1M PBS with 1% Triton X
Specification
Primary astrocyte culture
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Methanol
Username

Dr. Ruma Raha-Chowdhury

Verified customer

提交于 Sep 06 2016

Application
Western blot
Sample
Rabbit Tissue lysate - whole (Synovial membrane)
Gel Running Conditions
Reduced Denaturing (8 %)
Loading amount
35 µg
Specification
Synovial membrane
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jun 13 2016

Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
Yes - BD PERMEABILISATION SOLUTION
Sample
Mouse Cell (liver cells)
Specification
liver cells
Gating Strategy
CD24 POSITIVE
Preparation
Cell harvesting/tissue preparation method: single cell suspension
Sample buffer: 0.1%PBS-BSA
Username

Abcam user community

Verified customer

提交于 Dec 10 2013

Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
Yes - BD Permeabilisation Solution
Sample
Rat Cell (Liver cells)
Specification
Liver cells
Gating Strategy
CD24 Positive
Preparation
Cell harvesting/tissue preparation method: Single cell suspension
Sample buffer: 0.1% PBS-BSA
Username

Abcam user community

Verified customer

提交于 Dec 05 2013

Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
Yes - BD PERMEABILISATION SOLUTION
Sample
Human Cell (HEP G2)
Specification
HEP G2
Gating Strategy
CD24 POSITIVE
Preparation
Cell harvesting/tissue preparation method: single cell suspension
Sample buffer: 0.1%PBS-BSA
Username

Mr. brahmi manali

Verified customer

提交于 Dec 04 2013

Application
Western blot
Sample
Pig Tissue lysate - whole (small intestine)
Loading amount
30 µg
Specification
small intestine
Gel Running Conditions
Reduced Denaturing (7)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6% · Temperature: 20°C
Username

Mr. Dongil Kim

Verified customer

提交于 Apr 23 2013


Yes, I can send you as a one time exception a positive control lysate (rat liver (ab29413), http://www.abcam.com/liver-rat-whole-cell-lysate-normal-tissue-ab29413.html). This has been tested with ab66217 and gave strong bands.

Also I woul...

Read More

In this image the lower band is actin which has been recognized by and anti-actin antibody. This has been used as a loading control to normalize the levels of protein detected by confirming that protein loading is the same across the gel. The expressio...

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

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