PA5-25804

antibody from Invitrogen Antibodies

Targeting: CLTCL1 CHC22, CLH22, CLTCL, CLTD

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-25804

Provider

Invitrogen Antibodies

Product name

Clathrin Heavy Chain Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with bovine, mouse and rat based on sequence homology.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

400 µL

Concentration

0.25 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Clathrin Heavy Chain in Hela whole cell lysate. Samples were incubated with Clathrin Heavy Chain polyclonal antibody (Product # PA5-25804) using a dilution of 1:1,000 followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at a dilution of 1:10,000. Lysates/proteins: 20 µg per lane. Predicted band size: 192 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Clathrin Heavy Chain in formalin fixed and paraffin embedded human brain tissue. Samples were incubated with Clathrin Heavy Chain polyclonal antibody (Product # PA5-25804) followed by peroxidase conjugation of the secondary antibody and DAB staining. This data demonstrates the use of this antibody for immunohistochemistry. Clinical relevance has not been evaluated.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Clathrin Heavy Chain in formalin fixed and paraffin embedded human brain tissue. Samples were incubated with Clathrin Heavy Chain polyclonal antibody (Product # PA5-25804) followed by peroxidase conjugation of the secondary antibody and DAB staining. This data demonstrates the use of this antibody for immunohistochemistry. Clinical relevance has not been evaluated.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of HeLa cells using a CLTC polyclonal antibody (Product # PA5-25804) (right) compared to a negative control cell (left) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of Clathrin Heavy Chain in Hela cells (right histogram). Samples were incubated with Clathrin Heavy Chain polyclonal antibody (Product # PA5-25804) followed by FITC-conjugated goat-anti-rabbit secondary antibody. Negative control cell (left histogram).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1 W13-induced PM tubules provide an internalization pathway for beta1-integrin. ( a ) COS1 cells grown on coverslips expressing the membrane marker GFP-mem were incubated with an anti-beta1-integrin rat antibody for 30 minutes at 4 degC to avoid endocytosis, followed by incubation for 10 minutes at 37 degC to allow internalization in the presence of W13 (20 min, 4.5 ug/ml). After fixation, beta1-integrin was detected with an Alexa-555 labeled anti-rat antibody, and images were acquired with a confocal microscope (Leica TCS SP5). The higher magnification insets show beta1-integrin localization in W13-induced tubules (green arrowheads). ( b , c ) Following the same procedure explained as in ( a ), beta1-integrin was detected with an Alexa-647 labeled anti-rat antibody and EEA1 with a specific antibody and the secondary Alexa-555 anti-mouse in untreated ( b ) or W13-treated cells ( c ). Insets show beta1-integrin in EEA1-positive endosomes (red arrowheads) ( b ) or in tubules (green arrowheads) ( c ) (bars, 10 um). ( d , e ) Quantification of internalized beta1-integrin ( d ) and transferrin ( e ), as explained in the Materials and Methods , in COS1 cells expressing GFP-mem or GFP-Rac1 G12V for the indicated conditions (W13t, cells presenting tubules; W13nt, cells without tubules). Mean values +- standard error of the mean (SEM) from three independent experiments are shown. Statistical significance between different conditions and the corresponding control was determined