Anti-Clathrin heavy chain antibody (ab21679)

Rabbit polyclonal Clathrin heavy chain antibody. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Hamster, Human. Cited in 59 publication(s). Independently reviewed in 33 review(s).

Overview

  • Product name

    Anti-Clathrin heavy chain antibody
    See all Clathrin heavy chain primary antibodies
  • Description

    Rabbit polyclonal to Clathrin heavy chain
  • Host species

    Rabbit
  • Tested applications

    Suitable 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.

    Read Abcam's proprietary immunogen policy (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. ICC/IF: HeLa cell line.

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

  • Function

    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.
  • Sequence similarities

    Belongs to the clathrin heavy chain family.
  • Cellular localization

    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
  • 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

Images

  • ab21679 staining Clathrin in HeLa cells. The cells were fixed with 10% formaldehyde (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab21679 at 1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • 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

    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
    All lanes : 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

This product has been referenced in:

See all 72 Publications for this product

Customer reviews and Q&As

1-10 of 44 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (C166 endothelial cells)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
30 µg
Treatment
1 µg/ml LPS for 1 h
Specification
C166 endothelial cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Barbara Thaler

Verified customer

Submitted Sep 03 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (HUVEC cells)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
50 µg
Treatment
1 µg/ml LPS for 1 h
Specification
HUVEC cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Barbara Thaler

Verified customer

Submitted Sep 03 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela)
Permeabilization
Yes - Triton X100
Specification
Hela
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 19 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (LLC-PK1 cells)
Permeabilization
Yes - 0.4% saponin
Specification
LLC-PK1 cells
Blocking step
10%FBS, 2%BSA in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 27 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Podocytes)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
10 µg
Specification
Mouse Podocytes
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 27 2018

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

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

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

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

Abcam user community

Verified customer

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

Ms. Ya Zhou

Verified customer

Submitted Oct 06 2015

1-10 of 44 Abreviews or Q&A

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