antibody

Macrophage Scavenger Receptor I/CD204, Affinity Purified Antibody, Rabbit

MBS555114

0.1 mL

380 USD

polyclonal

rabbit

nonconjugated

human, mouse, rat, cow

western blot, immunocytochemistry

Antibody

Macrophage Scavenger Receptor I/CD204, Affinity Purified Antibody, Rabbit

[Macrophage Scavenger Receptor I/CD204]

[macrophage scavenger receptor types I and II isoform type 2; Macrophage scavenger receptor types I and II; macrophage scavenger receptor types I and II; macrophage scavenger receptor 1; Macrophage acetylated LDL receptor I and II; Scavenger receptor class A member 1; CD_antigen: CD204]

[MSR1; MSR1; SRA; SR-A; CD204; phSR1; phSR2; SCARA1; SCARA1]

MSRE_HUMAN

polyclonal

rabbit

Human, Mouse, Rat, Cat, Primate, and Bovine

358

Liquid PBS, 30% glycerol internal with 0.1% sodium azide

1.19 mg/ml

Store frozen. Aliquot as undiluted antisera and immediately place at -20°C. Antisera may have become trapped in top of vial during shipping. Centrifugation of vial is recommended before opening. Stable for 6 months at -20°C. Repeated freeze/thaw cycles compromise the integrity of the antiserum.

Immunocytochemistry (ICC), Immunofluorescence (IF), Western Blot (WB)

Dilution: . Western Blot: 0.5 ug/ml. Immunocytochemistry: 1:50-1:200. Immunofluorescence: 1:50-1:200. Dilutions listed as a recommendation. Optimal dilution should be determined by investigator. Localization: . Membrane; Single-pass type II membrane protein. This antibody is useful for Western Blot, where a band is seen ~50 kDa. Western Blot Protocol: . 1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 40 ug of total protein per lane. 2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus. 3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil. 4. Rinse the blot in TBS for approximately 5 minutes. 5. Block the membrane using 5% NFDM + 1% BSA in TBS + Tween, 1 hour at RT. 6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each. 7. Dilute the rabbit anti-MSR1 primary antibody (NBP1-100092) in blocking buffer and incubate 1 hour at room temperature. 8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each. 9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature. 10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background). 11. Apply the detection reagent of choice in accordance with the manufacturers’ instructions (Pierce ECL). Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding. Immunocytochemistry Protocol: . Culture cells to appropriate density in 35mm culture dishes or 6-well plates. 1. Pull off culture medium with and add 10% formalin to the dish. Fix at room temperature for 30 minutes. 2. Take off the formalin and add ice cold methanol (kept in well sealed bottle in -20C). Incubate for 5-10 minutes. 3. Take off methanol and add PBS (You can add 0.1% Tween-20 to PBS used here and all subsequent steps), be sure to not let the specimen dry out. Wash 3 times 10 minutes before proceeding to blocking step. 4. To block nonspecific antibody binding incubate in 10% normal goat serum for a minimum of 1 hr at room temp. Cells can also block overnight at 4C for this step. 5. Add primary antibody at appropriate dilution and incubate at room temp for 2 hrs or overnight at room temp. 6. Remove primary antibody and replace with PBS. Wash 3 x 10 min in PBS. 7. Add secondary antibody at appropriate dilution. Incubate for 1 hr at room temperature. 8. Remove antibody and replace with PBS, wash 1 x 10 min in PBS. Add Hoechst 33258 to PBS at 1:25,0000 and incubate for 10 min. Wash a third time with PBS for 10 min (total of 3X10min PBS washes). 9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide and parafilmed. Cells can also be coverslipped using Fluoromount. If storing coverslip be sure to seal the edges with clear nail polish. The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures. Immunohistochemistry Protocol: . Frozen Sections- . 1. Snap frozen fresh tissues in liquid nitrogen or isopentane pre-cooled in liquid nitrogen, embedded in OCT compound in cryomolds. Store frozen blocks at -80°C. 2. Cut 4-8 um thick cryostat sections and mount on superfrost plus slides or gelatin coated slides. Store slides at – 80°C until needed. 3. Before staining, warm slides at room temperature for 30 minutes and fix in ice cold acetone for 10 minutes. Air dry for 30 minutes. 4. Wash in PBS. Paraffin Sections- . 1. Deparaffinize sections in xylene, 2x5min. 2. Hydrate with 100% ethanol, 2x3min. 3. Hydrate with 95% ethanol, 1min. 4. Rinse in distilled water. Tissue Staining- . 1. Follow procedure for pretreatment as required. 2. Procedure for Immunoenzyme Staining. 3. Rinse Sections in Washing Buffer (PBS+0.1% Triton X-100, pH 7.4) for 2x2 min. 4. Serum Blocking: incubate sections in normal serum block – species same as secondary antibody( Wash Buffer + 5% Normal Sera). Note: since this protocol uses avidin-biotin detection system, avidin/biotin block may be needed based on tissue type. If you do, the avidin/biotin blocking should be done after normal serum block and before primary antibody incubation. 5. Primary Antibody: incubate sections in primary antibody at appropriate dilution in Wash Buffer for 1 hour at room temperature or overnight at 4C. Note:Do not rinse sections between serum block and primary antibody incubation. 6. Rinse in washing buffer for 3x2 min. 7. Peroxidase Blocking: incubate sections in peroxidase blocking solution for 10 minutes at room temperature. 8. Rinse in washing buffer for 3x2 min. 9. Secondary Antibody: incubate sections in biotinylated secondary antibody at an appropriate dilution in Wash Buffer for 30 minutes at room temperature. 10. Rinse in washing buffer for 3x2 min. 11. Detection: incubate sections in streptavidin-HRP in Wash Buffer for 30 minutes at room temperature. 12. Rinse in washing buffer for 3x2 min. 13. Chromagen/Substrate: incubate sections in peroxidase substrate solution. 14. Rinse in washing buffer for 3x2 min. 15. Rinse in running tap water for 2-5 minutes. 16. Dehydrate through 95% ethanol for 1 minute, 100% ethanol for 2x3min. 17. Clear in xylene for 2x5min. 18. Coverslip with mounting medium (i-BRITE Plus-Catalog#:MBS555421). Peroxidase Block- . 0.3% H2O2 in PBS (for Paraffin Sections). 0.3% H2O2 in Methanol (for Frozen Sections). Avidin/Biotin Block- . 0.001% Avidin in PBS. 0.001% Biotin in PBS. Block for 10 minutes with first solution, rinse with PBS, block for 10 minutes with second solution. The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

Immunofluorescence

Western Blot

Immunogen Sequence: Synthetic peptide made to a portion of the human protein (within residues 300-400). [Swiss-Prot# P21757].

Antibodies; Neurodegenerative Disease; Macrophage Scavenger Receptor I/CD204

Macrophage scavenger receptors (MSR) are membrane glycoproteins that mediate the recognition and uptake of a variety of macromolecules, including modified lipoproteins, advanced glycation end (AGEs) products and amyloid b-protein (Abeta). While the normal role of MSR is associated with cell adhesion and host defense mechanisms, it also has been implicated in the development of atherosclerosis and Abeta deposition in Alzheimer's disease (AD).

4505259

NP_002436.1

NM_002445.3

42,942 Da

AGE/RAGE Pathway (698754); Binding And Uptake Of Ligands By Scavenger Receptors Pathway (1269897); Phagosome Pathway (153910); Phagosome Pathway (153859); Scavenging By Class A Receptors Pathway (1269899); Vesicle-mediated Transport Pathway (1269876)

This gene encodes the class A macrophage scavenger receptors, which include three different types (1, 2, 3) generated by alternative splicing of this gene. These receptors or isoforms are macrophage-specific trimeric integral membrane glycoproteins and have been implicated in many macrophage-associated physiological and pathological processes including atherosclerosis, Alzheimer's disease, and host defense. The isoforms type 1 and type 2 are functional receptors and are able to mediate the endocytosis of modified low density lipoproteins (LDLs). The isoform type 3 does not internalize modified LDL (acetyl-LDL) despite having the domain shown to mediate this function in the types 1 and 2 isoforms. It has an altered intracellular processing and is trapped within the endoplasmic reticulum, making it unable to perform endocytosis. The isoform type 3 can inhibit the function of isoforms type 1 and type 2 when co-expressed, indicating a dominant negative effect and suggesting a mechanism for regulation of scavenger receptor activity in macrophages. [provided by RefSeq, Jul 2008]

MSR1: Membrane glycoproteins implicated in the pathologic deposition of cholesterol in arterial walls during atherogenesis. Two types of receptor subunits exist. These receptors mediate the endocytosis of a diverse group of macromolecules, including modified low density lipoproteins (LDL). Isoform III does not internalize actetylated LDL. Defects in MSR1 may be a cause of prostate cancer (PC). A malignancy originating in tissues of the prostate. Most prostate cancers are adenocarcinomas that develop in the acini of the prostatic ducts. Other rare histopathologic types of prostate cancer that occur in approximately 5% of patients include small cell carcinoma, mucinous carcinoma, prostatic ductal carcinoma, transitional cell carcinoma, squamous cell carcinoma, basal cell carcinoma, adenoid cystic carcinoma (basaloid), signet- ring cell carcinoma and neuroendocrine carcinoma. MSR1 variants may play a role in susceptibility to prostate cancer. MSR1 variants have been found in individuals with prostate cancer and co-segregate with the disease in some families. Defects in MSR1 may be a cause of Barrett esophagus (BE). A condition characterized by a metaplastic change in which normal esophageal squamous epithelium is replaced by a columnar and intestinal-type epithelium. Patients with Barrett esophagus have an increased risk of esophageal adenocarcinoma. The main cause of Barrett esophagus is gastroesophageal reflux. The retrograde movement of acid and bile salts from the stomach into the esophagus causes prolonged injury to the esophageal epithelium and induces chronic esophagitis, which in turn is believed to trigger the pathologic changes. Genetic variants in MSR1 have been found in individuals with Barrett esophagus and are thought to contribute to disease susceptibility. 3 isoforms of the human protein are produced by alternative splicing. Protein type: Membrane protein, integral. Chromosomal Location of Human Ortholog: 8p22. Cellular Component: collagen; integral to plasma membrane; plasma membrane; cytosol. Molecular Function: protein binding; low-density lipoprotein binding; scavenger receptor activity. Biological Process: receptor-mediated endocytosis; cholesterol transport; lipoprotein transport. Disease: Prostate Cancer; Barrett Esophagus

0.1 mL

380 USD