Overview

  • Product nameAnti-Clathrin heavy chain antibody
    See all Clathrin heavy chain primary antibodies
  • Description
    Rabbit polyclonal to Clathrin heavy chain
  • Tested applicationsSuitable for: WB, ICC/IF, IHC-P, Flow Cyt, IHC-FoFr, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Hamster, Human
    Predicted to work with: Chicken
  • Immunogen

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

    (Peptide available as ab23440.)

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

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • FunctionClathrin 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.
  • Sequence similaritiesBelongs to the clathrin heavy chain family.
  • Cellular localizationCytoplasmic 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
  • Database links
  • Alternative names
    • 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

Anti-Clathrin heavy chain antibody images

  • 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

References for Anti-Clathrin heavy chain antibody (ab21679)

This product has been referenced in:
  • Chang HF  et al. Endocytosis of Cytotoxic Granules Is Essential for Multiple Killing of Target Cells by T Lymphocytes. J Immunol 197:2473-84 (2016). Read more (PubMed: 27527597) »
  • Valley CC  et al. Sequential superresolution imaging of multiple targets using a single fluorophore. PLoS One 10:e0123941 (2015). Read more (PubMed: 25860558) »

See all 31 Publications for this product

Product Wall

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

Mr. Ephrem Kassa

Verified customer

Submitted 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

Submitted 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

Submitted 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

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Submitted 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

Submitted 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

Submitted 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
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Abcam user community

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Submitted 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
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Submitted 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

Submitted 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

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Submitted Jul 14 2014

1-10 of 39 Abreviews or Q&A

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