Rabbit Anti Human MIP-1a polyclonal, antigen affinity

///Rabbit Anti Human MIP-1a polyclonal, antigen affinity

Rabbit Anti Human MIP-1a polyclonal, antigen affinity

$150.00$190.00


accession P10147


Applications

ELISA:This antibody can be used at 2 μg/mL jointly with biotinylated Human MIP-1a antibody to detect Human MIP-1a. The detection limit for recombinant Human MIP-1a 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 MIP-1a. The detection limit for recombinant Human MIP-1a is approximately 0.2 ng/lane.

Source This antibody was produced in Rabbit immunized with recombinant Human MIP-1a.
Species reactivity Human
Purification The specific antibody was purified from Rabbit sera by using immobilized recombinant Human MIP-1a 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 P32302
Interactor P32246
Interactor P51677
Interactor P51681
Biological Process Chemotaxis
Biological Process Inflammatory-response
Molecular function Cytokine

Methods

Immunohistochemistry

  • Formalin-fixed paraffin-embedded (FFPE) 5 μm sections were deparaffinised and rehydrated by passage through and graduated alcohols.
  • Antigen retrieval was via microwaving in pre-heated EDTA buffer (0.37 g EDTA in 1 l distilled water, pH adjusted to 8.0 using 1N sodium hydroxide, 0.05% Tween 20) for 15 min.
  • Frozen 5 μm sections were thawed and fixed in acetone for 5 min.
  • Endogenous peroxidase was blocked by incubation with Peroxidase-Blocking Solution for 10 min.
  • Fc receptors were blocked by incubation in 10% casein solution for 30 min.
  • Sections were incubated in a primary antibody solution to the target antigen at a pre-determined dilution (goat polyclonal anti-CCL3, anti-CCL4, anti-CCL5, anti-CCL20, all at 10 μg/ml& , , ) or an isotype-matched control antibody solution at equal concentration for 1 h at room temperature.
  • The sections were incubated in an HRP-conjugated development solution and visualised in either ImmPACT NovaRED or ImmPACT DAB-Nickel .
  • Sections were counter-stained in Meyer's haematoxylin , cleared and mounted in DPX .
  • Tissue expression was…
  • Formalin-fixed paraffin-embedded (FFPE) 5 μm sections were deparaffinised and rehydrated by passage through and graduated alcohols.
  • Antigen retrieval was via microwaving in pre-heated EDTA buffer (0.37 g EDTA in 1 l distilled water, pH adjusted to 8.0 using 1N sodium hydroxide, 0.05% Tween 20) for 15 min.
  • Frozen 5 μm sections were thawed and fixed in acetone for 5 min.
  • Endogenous peroxidase was blocked by incubation with Peroxidase-Blocking Solution for 10 min.
  • Fc receptors were blocked by incubation in 10% casein solution for 30 min.
  • Sections were incubated in a primary antibody solution to the target antigen at a pre-determined dilution (goat polyclonal anti-CCL3, anti-CCL4, anti-CCL5, anti-CCL20, all at 10 μg/ml& , , ) or an isotype-matched control antibody solution at equal concentration for 1 h at room temperature.
  • The sections were incubated in an HRP-conjugated development solution and visualised in either ImmPACT NovaRED or ImmPACT DAB-Nickel .
  • Sections were counter-stained in Meyer's haematoxylin , cleared and mounted in DPX .
  • Tissue expression was visualised using aDM6000 microscope and the manufacturer's software.

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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