Rabbit Anti Human IL-6 polyclonal, antigen affinity

///Rabbit Anti Human IL-6 polyclonal, antigen affinity

Rabbit Anti Human IL-6 polyclonal, antigen affinity

$150.00$190.00


accession P05231


Applications

ELISA:This antibody can be used at 2 μg/mL jointly with biotinylated Human IL-6 antibody to detect Human IL-6. The detection limit for recombinant Human IL-6 is approximately 0.2 ng/well.

Western Blot:This antibody can be used at 0.2 μg/mL with the appropriate secondary reagents to detect Human IL-6. The detection limit for recombinant Human IL-6 is approximately 0.2 ng/lane.

Source This antibody was produced in rabbits immunized with recombinant Human IL-6.
Species reactivity Human
Purification The specific antibody was purified from Rabbit sera by using immobilized recombinant Human IL-6 affinity chromatography.
Presentation Lyophilized from PBS, pH 7.2.
Storage The lyophilized antibody is stable for at least 1 year from date of receipt at -20° C. Upon reconstitution, this antibody can be stored in working aliquots at 2° - 8° C for one month, or at -20° C for 12 months without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage This antibody product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.


Interactor P01730 CD4_HUMAN
Interactor P08887 IL6RA_HUMAN
Interactor P08887 IL6RA_HUMAN
Interactor P13725 ONCM_HUMAN
Interactor P40189 IL6RB_HUMAN
Biological Process Acute-phase
Molecular function Cytokine
Molecular function Growth-factor

Methods

Immunohistochemistry and Immunofluorescence Microscopy

  • Human and mice joint tissues were sectioned at 5-µm thickness for immunohistochemical staining.
  • Antigen retrieval was performed by incubating sections with 0.1% trypsin for 40 min at 37°C or with citrate buffer for 20 min at 95°C.
  • The following primary antibodies were used for immunohistochemistry: rabbit anti–HIF-2α and rabbit anti-ANKL , rabbit anti-MMP3 and anti-MMP13 , goat anti-IL6 , rabbit anti-IL17A , mouse anti-HIF-1α , and rat anti-C31 (ianova).
  • For double-immunofluorescence labeling of human and mouse joint tissues, the following primary antibodies were used: rabbit anti–HIF-2α ( for human tissues and for mouse tissues), rabbit anti-MMP3 , mouse anti-MMP13 (for human synovia), rabbit anti-MMP13 (for mouse synovia), goat anti-IL6 , mouse anti-vimentin (B ), rabbit anti-C55 , rabbit anti-C68 (for human synovia), rat anti-C68 (for mouse synovia), rabbit anti-TAP , rabbit anti-VEGF , rabbit anti-IL17A , rabbit anti-Ki67 , and rabbit anti-ANKL .
  • Expression levels of HIF-2α in RA synovium were quantified using Image…
  • Human and mice joint tissues were sectioned at 5-µm thickness for immunohistochemical staining.
  • Antigen retrieval was performed by incubating sections with 0.1% trypsin for 40 min at 37°C or with citrate buffer for 20 min at 95°C.
  • The following primary antibodies were used for immunohistochemistry: rabbit anti–HIF-2α and rabbit anti-ANKL , rabbit anti-MMP3 and anti-MMP13 , goat anti-IL6 , rabbit anti-IL17A , mouse anti-HIF-1α , and rat anti-C31 (ianova).
  • For double-immunofluorescence labeling of human and mouse joint tissues, the following primary antibodies were used: rabbit anti–HIF-2α ( for human tissues and for mouse tissues), rabbit anti-MMP3 , mouse anti-MMP13 (for human synovia), rabbit anti-MMP13 (for mouse synovia), goat anti-IL6 , mouse anti-vimentin (B ), rabbit anti-C55 , rabbit anti-C68 (for human synovia), rat anti-C68 (for mouse synovia), rabbit anti-TAP , rabbit anti-VEGF , rabbit anti-IL17A , rabbit anti-Ki67 , and rabbit anti-ANKL .
  • Expression levels of HIF-2α in RA synovium were quantified using Image J software.
  • The percentage of cells expressing HIF-2α was analyzed in synovial lining cells (fibroblast-like and macrophage-like synoviocytes), sublining macrophages, and endothelial cells

Read more

Immunohistochemistry and Immunofluorescence Microscopy

  • Human and mice joint tissues were sectioned at 5-µm thickness for immunohistochemical staining.
  • Antigen retrieval was performed by incubating sections with 0.1% trypsin for 40 min at 37°C or with citrate buffer for 20 min at 95°C.
  • The following primary antibodies were used for immunohistochemistry: rabbit anti–HIF-2α and rabbit anti-ANKL , rabbit anti-MMP3 and anti-MMP13 , goat anti-IL6 , rabbit anti-IL17A , mouse anti-HIF-1α , and rat anti-C31 (ianova).
  • For double-immunofluorescence labeling of human and mouse joint tissues, the following primary antibodies were used: rabbit anti–HIF-2α ( for human tissues and for mouse tissues), rabbit anti-MMP3 , mouse anti-MMP13 (for human synovia), rabbit anti-MMP13 (for mouse synovia), goat anti-IL6 , mouse anti-vimentin (B ), rabbit anti-C55 , rabbit anti-C68 (for human synovia), rat anti-C68 (for mouse synovia), rabbit anti-TAP , rabbit anti-VEGF , rabbit anti-IL17A , rabbit anti-Ki67 , and rabbit anti-ANKL .
  • Expression levels of HIF-2α in RA synovium were quantified using Image…
  • Human and mice joint tissues were sectioned at 5-µm thickness for immunohistochemical staining.
  • Antigen retrieval was performed by incubating sections with 0.1% trypsin for 40 min at 37°C or with citrate buffer for 20 min at 95°C.
  • The following primary antibodies were used for immunohistochemistry: rabbit anti–HIF-2α and rabbit anti-ANKL , rabbit anti-MMP3 and anti-MMP13 , goat anti-IL6 , rabbit anti-IL17A , mouse anti-HIF-1α , and rat anti-C31 (ianova).
  • For double-immunofluorescence labeling of human and mouse joint tissues, the following primary antibodies were used: rabbit anti–HIF-2α ( for human tissues and for mouse tissues), rabbit anti-MMP3 , mouse anti-MMP13 (for human synovia), rabbit anti-MMP13 (for mouse synovia), goat anti-IL6 , mouse anti-vimentin (B ), rabbit anti-C55 , rabbit anti-C68 (for human synovia), rat anti-C68 (for mouse synovia), rabbit anti-TAP , rabbit anti-VEGF , rabbit anti-IL17A , rabbit anti-Ki67 , and rabbit anti-ANKL .
  • Expression levels of HIF-2α in RA synovium were quantified using Image J software.
  • The percentage of cells expressing HIF-2α was analyzed in synovial lining cells (fibroblast-like and macrophage-like synoviocytes), sublining macrophages, and endothelial cells

Read more

Immunohistochemistry and Immunofluorescence Microscopy

  • Human and mice joint tissues were sectioned at 5-µm thickness for immunohistochemical staining.
  • Antigen retrieval was performed by incubating sections with 0.1% trypsin for 40 min at 37°C or with citrate buffer for 20 min at 95°C.
  • The following primary antibodies were used for immunohistochemistry: rabbit anti–HIF-2α and rabbit anti-ANKL , rabbit anti-MMP3 and anti-MMP13 , goat anti-IL6 , rabbit anti-IL17A , mouse anti-HIF-1α , and rat anti-C31 (ianova).
  • For double-immunofluorescence labeling of human and mouse joint tissues, the following primary antibodies were used: rabbit anti–HIF-2α ( for human tissues and for mouse tissues), rabbit anti-MMP3 , mouse anti-MMP13 (for human synovia), rabbit anti-MMP13 (for mouse synovia), goat anti-IL6 , mouse anti-vimentin (B ), rabbit anti-C55 , rabbit anti-C68 (for human synovia), rat anti-C68 (for mouse synovia), rabbit anti-TAP , rabbit anti-VEGF , rabbit anti-IL17A , rabbit anti-Ki67 , and rabbit anti-ANKL .
  • Expression levels of HIF-2α in RA synovium were quantified using Image…
  • Human and mice joint tissues were sectioned at 5-µm thickness for immunohistochemical staining.
  • Antigen retrieval was performed by incubating sections with 0.1% trypsin for 40 min at 37°C or with citrate buffer for 20 min at 95°C.
  • The following primary antibodies were used for immunohistochemistry: rabbit anti–HIF-2α and rabbit anti-ANKL , rabbit anti-MMP3 and anti-MMP13 , goat anti-IL6 , rabbit anti-IL17A , mouse anti-HIF-1α , and rat anti-C31 (ianova).
  • For double-immunofluorescence labeling of human and mouse joint tissues, the following primary antibodies were used: rabbit anti–HIF-2α ( for human tissues and for mouse tissues), rabbit anti-MMP3 , mouse anti-MMP13 (for human synovia), rabbit anti-MMP13 (for mouse synovia), goat anti-IL6 , mouse anti-vimentin (B ), rabbit anti-C55 , rabbit anti-C68 (for human synovia), rat anti-C68 (for mouse synovia), rabbit anti-TAP , rabbit anti-VEGF , rabbit anti-IL17A , rabbit anti-Ki67 , and rabbit anti-ANKL .
  • Expression levels of HIF-2α in RA synovium were quantified using Image J software.
  • The percentage of cells expressing HIF-2α was analyzed in synovial lining cells (fibroblast-like and macrophage-like synoviocytes), sublining macrophages, and endothelial cells

Read more

Sandwich ELISA

  • 96-well maxisorp plates were coated with the capture antibody overnight, then blocked with 5% FCS in PBS for 1 hour, then washed.
  • Then, the protein standard was added, and plates were incubated for 2 hrs at 37°C.
  • Plates were washed and incubated with a biotinylated detection antibody for 1 hour at 37°C.
  • After washing, plates were incubated with HRP-conjugated streptavidin for 30 min at room temperature and washed again.
  • Plates were developed using the tetramethylbenzidine peroxidase substrate system and absorbance was measured at 450-615 nm using an automated plate reader.

Western Blot

  • The Recombinant immunogen was migrated by 12% SDS-PAGE and transferred to a 0.2 um PVDF membrane.
  • After blocking the membrane in blocking buffer (5% milk powder in 20 mM Tris-HCl pH 7.5, 500 mM NaCl, 0.1% (v/v) Tween 20 , the membrane was incubated with primary antibody ( at 4°C overnight.
  • Peroxidase-linked secondary anti-rabbit were used to detect the bound primary antibodies.
  • Enhanced chemiluminescence (ECL) reagents were used to visualize