Anti-Clathrin heavy chain抗体(ab21679)

概述

  • 产品名称
    Anti-Clathrin heavy chain抗体
    参阅全部 Clathrin heavy chain 一抗
  • 描述
    兔多克隆抗体to Clathrin heavy chain
  • 经测试应用
    适用于: WB, ICC/IF, IHC-P, Flow Cyt, IHC-FoFr, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Hamster, Human
    预测可用于: Chicken
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1650 to the C-terminus of Human Clathrin heavy chain.

    (Peptide available as ab23440.)

  • 阳性对照
    • This antibody gave a positive signal in HeLa, Jurkat, A431 & NIH3T3 cells, as well as human kidney tissues sections.

性能

应用

Our Abpromise guarantee covers the use of ab21679 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 180 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-FoFr Use at an assay dependent concentration. PubMed: 18852301
IP Use at an assay dependent concentration.

靶标

  • 功能
    Clathrin is the major protein of the polyhedral coat of coated pits and vesicles. Two different adapter protein complexes link the clathrin lattice either to the plasma membrane or to the trans-Golgi network.
  • 序列相似性
    Belongs to the clathrin heavy chain family.
  • 细胞定位
    Cytoplasmic vesicle membrane. Membrane > coated pit. Melanosome. Cytoplasmic face of coated pits and vesicles. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • 数据库链接
  • 别名
    • CHC17 antibody
    • Clathrin heavy chain antibody
    • Clathrin heavy chain 1 antibody
    • Clathrin heavy chain 2 antibody
    • Clathrin heavy chain like 1 antibody
    • Clathrin heavy chain on chromosome 17 antibody
    • Clathrin heavy polypeptide antibody
    • CLH 17 antibody
    • CLH 22 antibody
    • CLH-17 antibody
    • CLH1_HUMAN antibody
    • CLH17 antibody
    • CLH22 antibody
    • CLTC antibody
    • CLTCL antibody
    • CLTCL1 antibody
    • CLTCL2 antibody
    • CLTD antibody
    • Hc antibody
    • KIAA0034 antibody
    see all

图片

  • ICC/IF image of ab21679 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21679, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • Anti-Clathrin heavy chain antibody (ab21679) at 1/1000 dilution + HUVEC at 20 µg

    Secondary
    HRP conjugated goat anti-rabbit IgG at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 180 kDa


    Exposure time : 5 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Immunocytochemical analysis of Mouse bEnd.3 labeling Clathrin heavy chain with ab21679 at 1/300 dilution using secondary anti-rabbit-alexa fluor 488 at 1/500 dilution

    See Abreview

  • ab21679 staining Clathrin heavy chain in mouse embryonic fibroblasts by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde and permeabilized with -200C ethanol. Samples were incubated with primary antibody at 1/500 dilution (in 0.1%Saponin/1% BSA/PBS) for 1 hour. An Alexa Fluor®546-conjugated goat polyclonal to rabbit IgG (H+L) was used as secondary antibody at 1/500 dilution. Green color in image show positive staining with Alexa Fluor®546 conjugated secondary.

    See Abreview

  • All lanes : Anti-Clathrin heavy chain antibody (ab21679) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)
    Lane 4 : NIH 3T3 cytoplasmic extract lysate (ab14873)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG (Alexa Fluor® 680 at 1/15000 dilution

    Predicted band size : 180 kDa
    Observed band size : 180 kDa
  • Image courtesy of Human Protein Atlas

    ab21679 staining Clathrin heavy chain in human kidney. Paraffin embedded human (male) kidney tissue was incubated with ab21679 (1/7000 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab21679 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

  • ab21679 immunoprecipitating clathrin heavy chain from human HeLa S3 whole cell lysate (20ug). A protein A matrix was used with the antibody at a concentration of 4ug/mg of lysate. ab21679 was further used in the western blot stage at a dilution of 1/250. The image shows:

    Lane 1: Clathrin IP (ab21679)

    Lane 2: Control IP

    Lane 3: Input (10%)

    See Abreview

  • ab21679 immunoprecipitating clathrin heavy chain from rat PC12 whole cell lysate (20ug). A protein A matrix was used with the antibody at a concentration of 4ug/mg of lysate. ab21679 was further used in the western blot stage at a dilution of 1/250. The image shows:

    Lane 1: Clathrin IP (ab21679)

    Lane 2: Control IP

    Lane 3: Input (10%)

    See Abreview

  • ab21679 staining Clathrin heavy chain in mouse embryonic fibroblast cells by Flow Cytometery. Samples were prepared by homogenization and centrifugation in trypsin and they were washed in PBS. Cells were fixed in 70% 40C ethanol and cell population was gated on Non-junk by FSC/SSC. The primary antibody was diluted 1/2000 in 1% BSA/PBS and incubated with sample for 1 hour at 22°C. An Alexa Fuor 488 conjugated goat polyclonal to rabbit IgG (H&L) was used as secondary antibody at dilution at 1/500.

    See Abreview

文献

This product has been referenced in:
  • Poulsen ET  et al. An Aberrant Phosphorylation of Amyloid Precursor Protein Tyrosine Regulates Its Trafficking and the Binding to the Clathrin Endocytic Complex in Neural Stem Cells of Alzheimer's Disease Patients. Front Mol Neurosci 10:59 (2017). ICC/IF . Read more (PubMed: 28360834) »
  • Halamoda-Kenzaoui B  et al. The agglomeration state of nanoparticles can influence the mechanism of their cellular internalisation. J Nanobiotechnology 15:48 (2017). WB ; Human . Read more (PubMed: 28651541) »

See all 48 Publications for this product

客户评价及客户问答

Application
Immunocytochemistry/ Immunofluorescence
Sample
Dog Cell (Kidney, Proximal tubule)
Permeabilization
Yes - 0.1% saponin
Specification
Kidney, Proximal tubule
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Ephrem Kassa

Verified customer

提交于 Jul 04 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (pericytes)
Permeabilization
Yes - 0.1% triton in PBS
Specification
pericytes
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Dec 18 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Hippocampal neuron)
Permeabilization
Yes - 0.2%Triton
Specification
Hippocampal neuron
Blocking step
Serum as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Nov 30 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (neuron)
Permeabilization
Yes - 0.2%Triton
Specification
neuron
Blocking step
Serum as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Oct 13 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (COS-7)
Permeabilization
Yes - 0.2% triton
Specification
COS-7
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Formaldehyde
Username

Ms. Ya Zhou

Verified customer

提交于 Oct 06 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (Kidney)
Permeabilization
No
Specification
Kidney
Blocking step
3% BSA and 0.5% TX100 as blocking agent for 45 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
3.2% PFA and 0.5% GA
Username

Dr. Aaron Halpen

Verified customer

提交于 Sep 29 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Sample
Pig Cell (PEAC)
Specification
PEAC
Permeabilization
Yes - 0.1% Triton X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 27 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Chinese Hamster Cell lysate - whole cell (CHO)
Specification
CHO
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Jul 28 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Sample
Mouse Cell (bEnd.3)
Specification
bEnd.3
Permeabilization
Yes - PBT (PBS+0.1%Triton)
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 14 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Sample
Human Cell (HUVEC)
Specification
HUVEC
Permeabilization
Yes - PBT (PBS+0.1%Triton)
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 14 2014

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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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