Detection of Mouse Histone H3 by IHC-IF. Sample: FFPE section of Mouse colon carcinoma. Antibody: Affinity purified rabbit anti-Histone H3 (Cat. No. IHC-00465) used at a dilution of 1:100. Detection: Red-fluorescent goat anti-rabbit IgG highly cross-adsorbed Antibody Hilyte Plus 555 (A120-501E) used at a dilution of 1:100. Epitope Retrieval Buffer-High pH (IHC-101J) was substituted for Epitope Retrieval Buffer-Reduced pH.
Detection of Human KARS by IHC. Sample: FFPE section of human breast carcinoma. Antibody: Affinity purified rabbit anti-KARS (Cat. No. IHC-00471) used at a dilution of 1:100. Detection: Red-fluorescent goat anti-rabbit IgG highly cross-adsorbed Antibody Hilyte Plus 555 (A120-501E) used at a dilution of 1:100. Epitope Retrieval Buffer-High pH (IHC-101J) was substituted for Epitope Retrieval Buffer-Reduced pH.
Detection of Human Rad52 by IHC. Sample: FFPE section of human prostate carcinoma. Antibody: Affinity purified rabbit anti-Rad52 (Cat. No. IHC-00462) used at a dilution of 1:250. Detection: DAB staining using Immunohistochemistry Accessory Kit (Cat. No. IHC-101). Epitope Retrieval Buffer-High pH (IHC-101J) was substituted for Epitope Retrieval Buffer-Reduced pH.
Immunohistochemistry Accessory Kit (Cat. No. IHC-101) provides the reagents and method for sensitive, reliable and economical immunohistochemical staining. This Kit is designed for immunostaining formalin-fixed, paraffin-embedded tissue sections with primary antibodies made in rabbit. It contains all the reagents required for the immunostaining procedure and will stain 250 slides.
Detection of Human and Mouse Vinculin by WB (h&m) and IP (h). Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded), 293T (T; 50 mcg), and mouse NIH3T3 (M; 50 mcg) cells. Antibodies: Vinculin antibody A302-535A used for WB at 0.1 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 10 mcg/mg lysate. Vinculin was not immunoprecipitated by Vinculin antibody A302-534A, which recognizes an upstream epitope. Detection: Chemiluminescence with exposure times of 3 minutes (A) and 10 seconds (B).
Detection of Human and Mouse RACK1 by WB (h&m) and Ip (h). Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded), 293T (T; 50 mcg), and mouse NIH3T3 (M; 50 mcg) cells. Antibodies: RACK1 antibody A302-545A used for WB at 0.1 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 3 mcg/mg lysate. RACK1 was also immunoprecipitated by rabbit anti-RACK1 antibody BL9179, which recognizes a downstream epitope. Detection: Chemiluminescence with exposure times of 3 minutes (A) and 30 seconds (B).
Detection of Human CARMA1 by WB and IP. Samples: Whole cell lysate from Ramos (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded) and K562 (K; 50 mcg) cells. Antibodies: CARMA1 antibody A302-542A used for WB at 0.1 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 10 mcg/mg lysate. CARMA1 was also immunoprecipitated by rabbit anti-CARMA1 antibodies A302-541A, A302-543A and A302-544A, which recognize other epitopes. Detection: Chemiluminescence with exposure time of 30 seconds (A) and 10 seconds (B).
ReliaBLOT® IP/Western Blot Reagents and Procedures (patent pending) provide an improved method for the detection of immunoprecipitated proteins assayed via Western Blot. It seamlessly integrates into existing IP/WB Protocols with three simple changes. It is optimized for antibodies made in rabbit.
