Immunohistochemistry was performed on 4-um-thick paraffin-embedded tissue sections after pressure cooker antigen retrieval (Biocare Medical; 30 to 40 min at 122ºC) in citrate buffer (Dako Target Retrieval Solution S1699) using mouse monoclonal antibodies anti-Rb (clone G3-245, BD Biosciences, San Diego, CA; 1:100 dilution), anti-p16 (clone E6H4, MTM Labs, Heidelberg, Germany; 1:2 dilution), and anti-CD34 (clone QBEnd10; Dako, Carpinteria, CA; 1:400 dilution). Dako Envision+ Mouse (K4007) secondary antibody was used (30-min incubation at room temperature). The sections were developed using 3,30-diaminobenzidine as substrate and counterstained with Mayer's hematoxylin. Intact expression within the nuclei of endothelial and inflammatory cells served as internal positive controls for p16. Dako mouse IgG1 was used as a negative control. Nuclear staining for RB1 and p16 were scored by two of the authors (G.E. and A.M.E.) as negative (loss of expression) when <10% of cells showed nuclear staining.
Anti cd34 clone qbend10
Manufactured by Agilent Technologies
Sourced in United States
The Anti-CD34 (clone QBEnd10) is a monoclonal antibody that binds to the CD34 antigen, which is a cell surface glycoprotein expressed on hematopoietic stem and progenitor cells. This antibody is commonly used in laboratory applications for the identification and enumeration of CD34-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Anti rb clone g3 245, by BD (1 mentions)
Anti cd105 clone sn6h, by Agilent Technologies (1 mentions)
Envision flex hrp, by Agilent Technologies (1 mentions)
Pt link module, by Agilent Technologies (1 mentions)
Envision flex dab chromogen, by Agilent Technologies (1 mentions)
Proteinase k, by Agilent Technologies (1 mentions)
Anti cd31 clone jc70a, by Agilent Technologies (1 mentions)
Anti αsma antibody, by Novus Biologicals (1 mentions)
Alexa fluor 488 conjugated goat anti mouse igg antibody, by Thermo Fisher Scientific (1 mentions)
Bio safe coomassie blue, by Bio-Rad (1 mentions)
Anti cd68 clone pg m1, by Agilent Technologies (1 mentions)
3 protocols using anti cd34 clone qbend10
1
Immunohistochemistry for RB1 and p16 in Tissue
2
Immunohistochemical Analysis of Vascular Markers
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Slices of 3 μm were analyzed. For CD31 and CD34 analysis, dewaxing, hydration, and thermal unmasking of the antigen were conducted with the DAKO PT Link module and with a high-pH solution (EnVisionTMFLEX Target Retrieval Solution, High pH, K8004). For CD105 analysis, which required enzymatic digestion to expose antigenic determinants, dewaxing, hydration, and unmasking were done manually, and proteinase K (DAKO, S3020) was used (5 min, room temperature). The following primary antibodies were used: anti-CD31 (clone JC70A, Ready-to-Use, DAKO IR610), anti-CD34 (clone QBEnd 10, Ready-to-Use, DAKO IR632), and anti-CD105 (clone SN6h, 1:20, DAKO M3527). EnVisionTM FLEX/HRP (DAKO SM802), containing peroxidase and secondary antibodies were used for detection. Diaminobenzidine (EnVisionTM FLEX DAB+ Chromogen, DAKO DM827) was employed for secondary antibody visualization. Contrast staining with hematoxylin was employed.
3
Immunohistochemical Staining of Vascular Markers
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Anti-CD31 rabbit polyclonal antibody was obtained from Spring Biosciences (Catalogue number E11114; Pleasanton, CA, USA). Anti-αSMA antibody was purchased from Novus Biologicals (Clone SPM332; Littleton, CO, USA). Anti-CD68 (Clone PG-M1) and anti-CD34 (Clone QBEnd10) antibodies were obtained from DAKO (Carpinteria, CA, USA). Anti-fibrinogen β-chain and anti-plasminogen antibodies were from Sigma-Aldrich. Alexa Fluor 488-conjugated goat anti-mouse IgG antibody and Alexa Fluor 567-conjugated goat anti-rabbit IgG antibody were purchased from Life Technologies Japan. Bio-Safe Coomassie Blue was purchased from Bio-Rad Laboratories.
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