Secondary Antibodies and Conjugates

Transcription

1 biodescriptor biomolecules Full image -- Remove color from text box when placing image Secondary Antibodies and Conjugates Initiative Descriptor Primary Cover Header Popular Reagents for Quality Secondary Detection Cover Header

2 biomolecules Secondary Antibodies and Conjugates Popular Reagents for Quality Detection Popular Reagents for Quality Detection Sigma offers a wide selection of secondary antibodies and conjugates. This brochure presents some of our most popular secondary antibodies. Features High Sensitivity Species-specific options Lot-to-lot consistency Species Mouse Rabbit Goat Others, including rat, chicken, human Labels Enzymes (Peroxidase, Alkaline ) Fluorochrome (FITC, TRITC, R-PE, Cy 3, APC, CF and Atto) Others (Biotin, Gold) Applications WB, IF, IHC, ELISA, Flow Glial cells cultured from rat amygdala. Nuclei stained blue with DAPI and glial processes stained green with Rabbit Anti-GFAP (Cat. No. G9269) and Goat Anti-Rabbit IgG-FITC (Cat. No. F9887). From R. Gabr, West Virginia University, Dept of Pharmocology and Toxicology, Morgantown, WV. Introduction Secondary antibodies are host antibodies that bind to primary antibodies or antibody fragments. They are typically labeled with probes that make them useful for detection, purification or sorting applications. Secondary antibodies may be polyclonal antibodies or monoclonal antibodies. Polyclonal antibodies lack the specificity of monoclonal antibodies but frequently have higher sensitivities because polyclonal antibodies are a mixture of antibodies that may include very high affinity antibodies. Cloning may not effectively select the highest affinity antibody from a polyclonal host. Secondary antibodies are available with specificity for whole Ig molecules or antibody fragments such as the Fc or Fab regions. Sigma s polyclonal secondary antibodies are produced from the serum of host animals including, but not limited to, mouse, rabbit, goat, and sheep. Monoclonal secondary antibodies are produced from mouse hybridoma clones. Secondary antibodies are used in immunodetection, and immunoaffinity purification applications. Immunodetection applications include enzyme linked immunosorbent assays (ELISA); Western blotting, immunohistology, immunoblotting, immunostaining and cell based assays, and high throughput cell-based screening assays (HTS). Also, secondary antibodies are useful for fluorescence activated cell sorting or FACS. Sigma s commitment to manufacture and offer high quality antibodies at a competitive price is only the beginning. Sigma recognizes the importance of an ongoing partnership between the manufacturer and the customer. In the Sigma tradition, we will continue to emphasize service and provide all possible assistance to you: through customer service, technical service, and our award winning online tools, such as the Antibody Explorer and Your Favorite Gene; all of which allow you to search and browse for antibodies in a wide variety of ways. Jurkat cell nuclear extracts were separated on SDS- PAGE and blotted with Monoclonal Anti-MECP2, (lane 1) clone Men-8 (Cat. No. M7443); (lane 2) clone Mec-168 (Cat. No. M6818). The antibody was developed with Goat Anti-Mouse IgG (Fab specific)-peroxidase (Cat No. A2304) and a chemiluminescent substrate. For our complete listing of secondary antibodies and conjugates, visit sigma.com/secondaryabs

3 72 55 Order Technical Service The Ideal Anti-Rabbit Secondary Antibody for Western Blotting Sigma s Mouse Anti-Rabbit IgG, Native, clone RabT-50 does not recognize denatured rabbit IgG. This feature is extremely useful in applications where the immuno pre ci pi ta ting and detecting antibodies are both produced in rabbit. In the example, this function is specifically demonstrated in the negative control lanes. Using Anti-Rabbit IgG, Native (Cat. No. R2655) the negative control lanes are clear in Panel A. However in Panel B, if a traditional Anti-Rabbit secondary is applied, the heavy chain of the immunoprecipitating antibody interferes and masks the sample bands. This antibody is especially valuable when the protein of interest has a molecular mass around 50 kda, which can be masked by the denatured (heavy chain) IgG. B A Immunoprecipitation Rabbit Anti-eRF1 (Cat. No. E8156) was used to immunoprecipitate erf1 from HeLa cell lysate. (Lane 1) whole antiserum; (Lane 2) 5 mg; (Lane 3) 2.5 mg; (Lane 4) Negative control without cell lysate; (Lane 5) Negative control without immunoprecipitating antibody; (Lane 6) Immunoblotting positive control extract of HeLa probed with Rabbit Anti-eRF1 (Cat. No. E8156). Panel A. Detection secondary antibody Monoclonal Mouse Anti-Rabbit IgG, clone RabT-50 (Cat. No. R2655) followed by Goat Anti-Mouse IgG, Peroxidase Conjugate (Cat. No. A2304). Panel B. Detection secondary antibody Goat Anti-Rabbit IgG, Peroxidase Conjugate (Cat. No. A0545) B Monoclonal Secondaries Cat. 72 No. Product Description Host Clone Form Application A9452-1VL Monoclonal Anti-Goat/Sheep IgG Peroxidase mouse GT-34 purified immunoglobulin DB, ELISA (d) IHC (p), WB CL 55 R UL Monoclonal Anti-Rabbit IgG, Native mouse RabT-50 purified immunoglobulin ELISA (i), ICC, WB R UL 43 R UL Monoclonal Anti-Rabbit IgG, Native Peroxidase mouse RabT-50 purified immunoglobulin ELISA (i), ICC, WB A1949-1VL 34 Monoclonal Anti-Rabbit IgG (γ-chain specific) Peroxidase mouse RG-96 purified immunoglobulin DB, ELISA (d), IHC (p) A2074-1VL 26 Monoclonal Anti-Rabbit Immunoglobulins Peroxidase mouse RG-16 purified immunoglobulin ELISA (d), IHC (p), WB CL

4 biomolecules Goat Secondary Antibodies and Conjugates Cat. No. Product Description Host Form Application A ML Anti-Goat IgG (whole molecule) Alkaline rabbit affinity isolated antibody ELISA (d), IHC (p), WB A ML A4187-1ML A5420-1ML Anti-Goat IgG (whole molecule) Peroxidase rabbit affinity isolated antibody ELISA (d), IHC (p), WB CL SAB UL SAB UL SAB UL SAB UL SAB UL SAB UL SAB UL SAB UL SAB UL SAB UL SAB UL SAB UL Anti-Goat IgG (H+L), highly cross-adsorbed, donkey affinity isolated antibody FCM Biotin antibody Anti-Goat IgG (H+L), CF 488A antibody chicken affinity isolated antibody FCM Anti-Goat IgG (H+L), CF 594 antibody rabbit affinity isolated antibody FCM Anti-Goat IgG (H+L), CF 680 antibody rabbit affinity isolated antibody FCM Anti-Goat IgG (H+L), highly cross-adsorbed, CF 350 antibody Anti-Goat IgG (H+L), highly cross-adsorbed, CF 647 antibody donkey affinity isolated antibody FCM donkey affinity isolated antibody FCM Immunofluorescence Immunofluorescence Human HeLa cells were fixed and permeabilized with 4% paraformaldehyde followed by 0.5% Triton X-100. Fixed cells were stained with 10 µg/ml Anti-Nucleolin (Cat. No. N2662) and developed with Goat Anti-Rabbit IgG, FITC conjugate (Cat. No. F9887). Fluorescent micrographs of mitotic newt lung cells. Microtubules are stained with Monoclonal Anti-β-Tubulin and Goat Anti-Mouse IgG FITC conjugate (Cat. No. F0257). Chromosomes are counterstained with Hoechst The astral microtubules are green, the spindle is yellow, and the chromosomes are blue. From C. Rieder, Wadsworth Center, Division of Molecular Medicine, Lab of Cell Regulation, Albany, NY. MW 35 Annexin V Immunoblotting HeLa cell extract was separated by SDS-PAGE and probed with or without Monoclonal Anti-Annexin V, Clone AN5 (Cat. No. A8604) followed by Goat Anti-Mouse, Alkaline conjugate (Cat. No. A2179). A B MW 220 kda Immunoblotting Rat brain cortex whole cell extracts were separated on SDS-PAGE, blotted with Monoclonal Anti-MAP2 (2A+2B) (Cat. No. M1406) and developed using Goat Anti-Mouse IgG, Alkaline (Cat. No. A1293) and NBT/BCIP substrate. (Lane A) Antibody dilution 1:500; (Lane B) Negative control (only secondary antibody).

5 Order Technical Service Mouse Secondary Antibodies and Conjugates Cat. No. Product Description Host Form Application M1397-1MG Anti-Mouse IgG (γ-chain specific) goat affinity isolated antibody ODD M1397-5X1MG M2650-1ML Anti-Mouse IgG (Fc specific) goat affinity isolated antibody QPA M4280-1ML Anti-Mouse IgG (Fc specific) goat affinity isolated antibody ELISA (i) M8642-1MG Anti-Mouse IgG (whole molecule) goat affinity isolated antibody ODD M8642-5X1MG M8645-1MG Anti-Mouse IgG (whole molecule) goat affinity isolated antibody ODD M8645-5X1MG M7023-2ML Anti-Mouse IgG (whole molecule) rabbit IgG fraction of antiserum QPA A ML Anti-Mouse IgG (γ-chain specific) Alkaline goat affinity isolated antibody ELISA (d), IHC (p), WB A ML A3438-1ML A ML A ML A2429-1ML Anti-Mouse IgG (Fc specific) Alkaline goat affinity isolated antibody ELISA (d), IHC (p), WB CL A ML A ML A3562-1ML A5153-1ML F ML F0257-1ML F0257-2ML F0257-5X2ML F ML F2012-1ML F2012-2ML F ML F9137-1ML F9137-2ML Anti-Mouse IgG (whole molecule) Alkaline goat affinity isolated antibody ELISA (d), IHC (p), WB Anti-Mouse IgG (whole molecule) Alkaline goat affinity isolated antibody ELISA (d) Anti-Mouse IgG (whole molecule) FITC goat affinity isolated antibody IF (d) Anti-Mouse IgG (whole molecule) FITC goat affinity isolated antibody IF (d) Anti-Mouse IgG (whole molecule) FITC rabbit IgG fraction of antiserum IF (d), IHC (p) A3673-1ML Anti-Mouse IgG (γ-chain specific) Peroxidase goat affinity isolated antibody ELISA (d) A9917-1ML Anti-Mouse IgG (Fab specific) Peroxidase goat affinity isolated antibody ELISA (d), IHC (p), WB CL A2304-1ML Anti-Mouse IgG (Fab specific) Peroxidase goat affinity isolated antibody WB, ELISA (d), IHC (p) A0168-1ML Anti-Mouse IgG (Fc specific) Peroxidase goat affinity isolated antibody ICC, ELISA (d), IHC (p), WB CL A2554-1ML Anti-Mouse IgG (Fc specific) Peroxidase goat affinity isolated antibody DB, ELISA (d), IHC (p), WB CL A ML Anti-Mouse IgG (whole molecule) Peroxidase goat affinity isolated antibody ELISA (d) A4416-1ML A4416-5X1ML A ML Anti-Mouse IgG (whole molecule) Peroxidase goat affinity isolated antibody ELISA (d) A9044-2ML Anti-Mouse IgG (whole molecule) Peroxidase rabbit IgG fraction of antiserum ELISA (d), IHC (p), WB CL P ML Anti-Mouse IgG (whole molecule) F(ab ) 2 sheep affinity isolated antibody IF (i) fragment R-Phycoerythrin SAB UL Anti-Mouse IgG (H+L), CF 350 antibody goat affinity isolated antibody FCM, ICC, IHC, IF (i) SAB UL SAB UL SAB UL Anti-Mouse IgG (H+L), CF 750 antibody goat affinity isolated antibody FCM, ICC, IHC, IF (i)

6 biomolecules Rabbit Secondary Antibodies and Conjugates Cat. No. Product Description Host Form Application R2004-1MG Anti-Rabbit IgG (whole molecule) goat affinity isolated antibody IEP R2004-5X1MG A ML Anti-Rabbit IgG (whole molecule) Alkaline goat affinity isolated antibody ELISA (d), IHC (p), WB A ML A3687-1ML A ML A8025-1ML A8025-5X1ML Anti-Rabbit IgG (whole molecule) Alkaline goat affinity isolated antibody ELISA (d) A ML A ML A9919-1ML A ML A3812-1ML A ML A ML A3937-1ML Anti-Rabbit IgG (whole molecule) Alkaline goat affinity isolated antibody DB, ELISA (d), IHC (p), WB CL Anti-Rabbit IgG (whole molecule) Alkaline goat affinity isolated antibody ELISA (d), IHC (p), WB Anti-Rabbit IgG (whole molecule), F(ab ) 2 fragment Alkaline goat affinity isolated antibody ELISA (d), IHC (p), WB C2306-1ML Anti-Rabbit IgG (whole molecule), F(ab ) 2 fragment Cy3 sheep affinity isolated antibody IHC (p) F ML Anti-Rabbit IgG (whole molecule) FITC goat affinity isolated antibody IF (i) F0382-1ML F0382-2ML F ML Anti-Rabbit IgG (whole molecule) FITC goat affinity isolated antibody IF (i), IHC (p) F9887-1ML F9887-2ML A6154-1ML Anti-Rabbit IgG (whole molecule) Peroxidase goat affinity isolated antibody ELISA (d) A6154-5X1ML A0545-1ML Anti-Rabbit IgG (whole molecule) Peroxidase goat affinity isolated antibody ELISA (d), IHC (p), WB CL A4914-1ML Anti-Rabbit IgG (whole molecule) Peroxidase goat affinity isolated antibody ELISA (d) A9169-2ML Anti-Rabbit IgG (whole molecule) Peroxidase goat IgG fraction of antiserum ELISA (d), IHC (p), WB CL T6778-1ML Anti-Rabbit IgG (whole molecule) TRITC goat affinity isolated antibody IF (i), IHC (p) SAB UL Anti-Rabbit IgG (H+L), CF 405M antibody goat affinity isolated antibody FCM, ICC, IHC, IF (i) SAB UL SAB UL Anti-Rabbit IgG (H+L), CF 555 antibody goat affinity isolated antibody FCM, ICC, IHC, IF (i) SAB UL SAB UL Anti-Rabbit IgG (H+L), CF 660R antibody goat affinity isolated antibody FCM, ICC, IHC, IF (i) SAB UL SAB UL Anti-Rabbit IgG (H+L), highly cross-adsorbed, CF 350 antibody donkey affinity isolated antibody FCM, ICC, IHC, IF (i) SAB UL SAB UL Anti-Rabbit IgG (H+L), highly cross-adsorbed, CF 633 antibody goat affinity isolated antibody FCM, ICC, IHC, IF (i) SAB UL SAB UL SAB UL Anti-Rabbit IgG (H+L), highly cross-adsorbed, CF 647 antibody goat affinity isolated antibody FCM, ICC, IHC, IF (i) Immunofluorescence Primary neuronal culture of rat amygdala. Nuclei stained blue with DAPI (Cat. No. D9542), neuron cluster (lower left) ensheathed by glil processes stained red with Rabbit Anti-GFAP (Cat. No. G9269) and Goat Anti-Rabbit IgG-TRITC (Cat. No. T6778) and positive neuronal fibers stained green with Rabbit Anti-Neurofilament 200 (Cat. No. N4142) and Goat Anti-Rabbit IgG-FITC (Cat. No. F9887). From R. Gabr, West Virginia Univ., Dept of Pharmacology & Toxicology, Morgantown, WV. Immunofluorescence Plasma Membrane Caveoli Staining: HUVEC cells were fixed with 4% paraformaldehyde/0.1% Triton X-100 and stained with Anti-Caveolin-1 (Cat. No. C4490) at a 1:100 dilution, followed by Goat Anti-Rabbit IgG-FITC (Cat. No. F9887)

7 50,000 antibodies, 100% Bioguaranteed. Sigma Life Science has the antibody for your research needs. Bioguarantee. Sigma Life Science offers more than 50,000 highly validated primary and secondary antibodies easily accessed by our innovative Antibody Explorer online search tool, supported by our renowned technical service and backed by our industry leading, no hassle Bioguarantee.* Find your antibody: sigma.com/antibodyexplorer *Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase. All required fields of the Antibody Bioguarantee Form must be completed. Refunds and replacements contingent to claim review by technical service team. Credit covers the cost of antibody. Product replacements depend on product availability Sigma-Aldrich Co. LLC. All rights reserved. SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co. LLC, registered in the US and other countries. Where bio begins is a trademark of Sigma-Aldrich Co. LLC.

8 Sigma-Aldrich Worldwide Offices Argentina Free Tel: Tel: (+54) Fax: (+54) Australia Free Tel: Free Fax: Tel: (+61) Fax: (+61) Austria Tel: (+43) Fax: (+43) Belgium Tel: (+32) Fax: (+32) Brazil Free Tel: Tel: (+55) Fax: (+55) Canada Free Tel: Free Fax: Tel: (+1) Fax: (+1) Chile Tel: (+56) Fax: (+56) People s Republic of China Free Tel: Tel: (+86) Fax: (+86) Germany Free Tel: Free Fax: Tel: (+49) Fax: (+49) Hungary Tel: (+36) Fax: (+36) India Telephone Bangalore: (+91) New Delhi: (+91) Mumbai: (+91) Pune: (+91) Hyderabad: (+91) Kolkata: (+91) Fax Bangalore: (+91) New Delhi: (+91) Mumbai: (+91) Pune: (+91) Hyderabad: (+91) Kolkata: (+91) Ireland Free Tel: Free Fax: Tel: +353 (0) Fax: (0) Israel Free Tel: Tel: (+972) Fax: (+972) Czech Republic Tel: (+420) Fax: (+420) Italy Free Tel: Tel: (+39) Fax: (+39) Denmark Tel: (+45) Fax: (+45) Japan Tel: (+81) Fax: (+81) Finland Tel: (+358) Fax: (+358) Korea Free Tel: (+82) Free Fax: (+82) Tel: (+82) Fax: (+82) France Free Tel: Free Fax: Tel: (+33) Fax: (+33) Enabling Science to Improve the Quality of Life Luxembourg Tel: (+32) Fax: (+32) Malaysia Tel: (+60) Fax: (+60) Mexico Free Tel: Free Fax: Tel: (+52) Fax: (+52) The Netherlands Tel: (+31) Fax: (+31) New Zealand Free Tel: Free Fax: Tel: (+61) Fax: (+61) Norway Tel: (+47) Fax: (+47) Poland Tel: (+48) Fax: (+48) Portugal Free Tel: Free Fax: Tel: (+351) Fax: (+351) Russia Tel: (+7) Fax: (+7) Spain Free Tel: Free Fax: Tel: (+34) Fax: (+34) Sweden Tel: (+46) Fax: (+46) Switzerland Free Tel: Free Fax: Tel: (+41) Fax: (+41) Thailand Tel: (+66) Fax: (+66) United Kingdom Free Tel: Free Fax: Tel: (+44) Fax: (+44) United States Toll-Free: Toll-Free Fax: Tel: (+1) Fax: (+1) Vietnam Tel: (+84) Fax: (+84) Internet sigma-aldrich.com Singapore Tel: (+65) Fax: (+65) Slovakia Tel: (+421) Fax: (+421) South Africa Free Tel: Free Fax: Tel: (+27) Fax: (+27) Order/Customer Service (800) Fax (800) Technical Service (800) sigma-aldrich.com/techservice Development/Custom Manufacturing Inquiries (800) Safety-related Information sigma-aldrich.com/safetycenter World Headquarters 3050 Spruce St. St. Louis, MO (314) sigma-aldrich.com 2012 Sigma-Aldrich Co. LLC. All rights reserved. SIGMA, SAFC and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co. LLC, registered in the US and other countries. Where bio begins and SIGMAFAST are trademarks of Sigma-Aldrich Co. LLC. Cy is a registered trademark of GE Healthcare. Triton is a registered trademark of Dow Chemical Co. Sigma brand products are sold through Sigma-Aldrich, Inc. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see product information on the Sigma-Aldrich website at and/or on the reverse side of the invoice or packing slip. LCF