Skip to main content Skip to navigation
Purified Mouse Anti-Clathrin Heavy Chain
Purified Mouse Anti-Clathrin Heavy Chain

Immunohistochemical analysis of Clathrin heavy chain expression on rat Rat cerebellum (Left Panel). Zinc-fixed paraffin embedded tissue was stained with Purified Mouse Anti-Clathrin Heavy Chain (Cat. No. 610499, 610500), and visualized with secondary staining. Figure was taken at 40X magnification.

Western blot analysis of Clathrin Heavy Chain on HeLa cell lysate (Center Panel). HeLa cell lysate was stained with Purified Mouse Anti-Clathrin Heavy Chain antibody in the following dilutions: 1:1000 (lane 1), 1:2000 (lane 2), 1:4000 (lane 3). Clathrin heavy chain was visualized with HRP Goat Anti-Mouse Ig (Cat. No. 554002), and is expressed as a 180 kDa band.  

Immunofluorescent staining of C6 cells (Right Panel).  Cells were seeded in a collagen coated 384 well imaging plate at ~ 6,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and Purified Mouse Anti-Clathrin Heavy Chain antibody.  The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen).  The image was taken on a Pathway 855 or 435 imager using a 20x objective.  This antibody also stained SH-SY5Y, SK-N-SH cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure).

Immunohistochemical analysis of Clathrin heavy chain expression on rat Rat cerebellum (Left Panel). Zinc-fixed paraffin embedded tissue was stained with Purified Mouse Anti-Clathrin Heavy Chain (Cat. No. 610499, 610500), and visualized with secondary staining. Figure was taken at 40X magnification.

Western blot analysis of Clathrin Heavy Chain on HeLa cell lysate (Center Panel). HeLa cell lysate was stained with Purified Mouse Anti-Clathrin Heavy Chain antibody in the following dilutions: 1:1000 (lane 1), 1:2000 (lane 2), 1:4000 (lane 3). Clathrin heavy chain was visualized with HRP Goat Anti-Mouse Ig (Cat. No. 554002), and is expressed as a 180 kDa band.  

Immunofluorescent staining of C6 cells (Right Panel).  Cells were seeded in a collagen coated 384 well imaging plate at ~ 6,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and Purified Mouse Anti-Clathrin Heavy Chain antibody.  The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen).  The image was taken on a Pathway 855 or 435 imager using a 20x objective.  This antibody also stained SH-SY5Y, SK-N-SH cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure).

製品詳細
Down Arrow Up Arrow


BD Transduction Laboratories™
Human (QC Testing), Mouse, Rat, Dog, Chicken (Tested in Development)
Mouse IgG1
Rat Clathrin Heavy Chain aa. 4-171
Western blot (Routinely Tested), Bioimaging, Immunofluorescence, Immunohistochemistry, Immunoprecipitation (Tested During Development)
180 kDa
250 µg/ml
AB_397866
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


推奨アッセイ手順

Immunofluorescent staining and bioimaging

Methanol Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample.

Triton-X 100 Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.

製品通知

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Triton is a trademark of the Dow Chemical Company.
  6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
610499 Rev. 1
抗体の詳細
Down Arrow Up Arrow
23/Clathrin Heavy Chain

Clathrin is the major protein component in the coat formed around pits and vesicles involved in receptor-mediated endocytosis. Clathrin forms a non-covalently bound triskelion structure composed of three heavy chains (192 kDa each) and three light chains (23-25 kDa each). Each leg of the triskelion structure contains one heavy and one light chain. The three heavy chains forming the triskelion structure are attached at their respective proximal ends like spokes on a wheel. Clathrin heavy chain is composed of a terminal globular domain, a distal segment containing several areas sensitive to enzymatic cleavage, and a proximal segment which contains a light chain binding site. The proximal and distal domains are connected by a joint segment at which there is a sharp bend in the heavy chains of fully-assembled triskelia. Although the calculated molecular weight is 192 kDa, clathrin heavy chain migrates at approximately 180 kDa.

610499 Rev. 1
フォーマットの詳細
Down Arrow Up Arrow
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
610499 Rev.1
引用&参考文献
Down Arrow Up Arrow

開発者向け参考資料 (5)

  1. Kalthoff C, Groos S, Kohl R, Mahrhold S, Ungewickell EJ. Clint: a novel clathrin-binding ENTH-domain protein at the Golgi. Mol Biol Cell. 2002; 13(11):4060-4073. (Clone-specific: Immunofluorescence, Western blot). 参考文献を見る
  2. Liu SH, Wong ML, Craik CS, Brodsky FM. Regulation of clathrin assembly and trimerization defined using recombinant triskelion hubs. Cell. 1995; 83(2):257-267. (Biology). 参考文献を見る
  3. Okamoto CT, Karam SM, Jeng YY, Forte JG, Goldenring JR. Identification of clathrin and clathrin adaptors on tubulovesicles of gastric acid secretory (oxyntic) cells. Am J Physiol. 1998; 274(1):C1017-C1029. (Clone-specific: Immunohistochemistry, Western blot). 参考文献を見る
  4. Padilla PI, Chang MJ, Pacheco-Rodriguez G, Adamik R, Moss J, Vaughan M. Interaction of FK506-binding protein 13 with brefeldin A-inhibited guanine nucleotide-exchange protein 1 (BIG1): effects of FK506. Proc Natl Acad Sci U S A. 2003; 100(5):2322-2327. (Clone-specific: Immunoprecipitation, Western blot). 参考文献を見る
  5. van Kerkhof P, Sachse M, Klumperman J, Strous GJ. Growth hormone receptor ubiquitination coincides with recruitment to clathrin-coated membrane domains. J Biol Chem. 2001; 276(6):3778-3784. (Clone-specific: Electron microscopy). 参考文献を見る
すべて表示する (5) 表示項目を減らす
610499 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.