Cd68 Antibody (OASA04073)

Catalog No: OASA04073 (Formerly GWB-Q01303)

Size:100UG

Staining of rat liver, with induced hepatocellular damage, with Mouse anti Rat CD68

Staining of rat peritoneal macrophages cells with Mouse anti Rat CD68:RPE. Permeabilised with Leucoperm (Fix & Perm) (BUF09)

Staining of rat peritoneal macrophages cells with Mouse anti Rat CD68:FITC following permeabilisation with Leucoperm (BUF09)

Staining of rat peritoneal macrophages with Mouse anti Rat CD68: Alexa Fluor 647 following permeabilisation with Leucoperm (BUF09)

Staining of rat peritoneal macrophages with Mouse anti Rat CD68: Alexa Fluor 488. following permeabilisation with Leucoperm (BUF09)

Product Info

• Predicted Species Reactivity: Rat
• Product Format: Liquid. Phosphate buffered saline
• Clonality: Monoclonal
• Clone: ED1
• Isotype: IgG1
• Host: Mouse
• Application: Flow cytometry|Immunofluorescence|Immunohistochemistry-Frozen|Immunohistochemistry-Paraffin|Immunoprecipitation|Western blot
• Additional Information: Fusion Partners: Spleen cells from immunised BALB/c mice were fused with cells of the SP2/0-Ag14 mouse myeloma cell line.
  Preparation: Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
  Preservative Stabilisers: 0.09% - Sodium Azide
  Approx Protein Conc: IgG concentration 0.5 mg/ml
  Buffer Solutions: Phosphate buffered saline pH7.4
• Reconstitution and Storage: -20°C
• Immunogen: Rat spleen cells
• Purification: Protein A Purified
• Predicted Homology Based on Immunogen Sequence: Rat
• Concentration: 0.5 mg/ml
• Specificity: CD68
• Application Info: Flow Cytometry(1): 1/50 - 1/100
  
• Application Data: Application #1: Staining of rat liver, with induced hepatocellular damage, with Mouse anti Rat CD68
  Application #2: Staining of rat peritoneal macrophages cells with Mouse anti Rat CD68:RPE . Permeabilised with Leucoperm (Fix & Perm) (BUF09)
  Application #3: Staining of rat peritoneal macrophages cells with Mouse anti Rat CD68:FITC following permeabilisation with Leucoperm (BUF09)
  Application #4: Staining of rat peritoneal macrophages with Mouse anti Rat CD68: Alexa Fluor 647 following permeabilisation with Leucoperm (BUF09)
  Application #5: Staining of rat peritoneal macrophages with Mouse anti Rat CD68: Alexa Fluor 488 . following permeabilisation with Leucoperm (BUF09)
• Protocol Information: Citation: 1: Campbell SJ, Perry VH, Pitossi FJ, Butchart AG, Chertoff M, Waters S, DempsterR, Anthony DC. Central nervous system injury triggers hepatic CC and CXCchemokine expression that is associated with leukocyte mobilization andrecruitment to both the central nervous system and the liver. Am J Pathol. 2005May;166(5):1487-97. PubMed PMID: 15855648; PubMed Central PMCID: PMC1606402.
  Species: Rat
  Experiment Name: 1. Identification of Leukocytes2. Immunocytochemistry for CCL-2
  Experiment Background: The administration of interleukin-1β to the brain induces hepatic CXC chemokine synthesis, which increases neutrophil levels in the blood, liver, and brain. Sandra et al. showed that such hepatic response is not restricted to the CXC chemokines. CCL-2, a CC chemokine, was released by the liver in response to a tumor necrosis factor (TNF)-α challenge to the brain and boosted monocyte levels. Furthermore, a clinically relevant compression injury to the spinal cord triggered hepatic chemokine expression of both types.
  Experimental Steps: 1. Identification of LeukocytesFrozen, 10-um-thick serial coronal sections were cut from tissue blocks, through the injection site in the brain, through a representative lobe of liver or in 20-um-thick parasagittal sections through the compression-injured spinal cord. Using immunohistochemistry, neutrophils were identified using the anti-neutrophil serum HB199, activated macrophages/Kupffer cells and recruited monocytes were identified using the anti-ED-1 serum (Oxford, UK). For each tissue section, four representative fields were chosen for quantitation and the average number of positive cells was calculated and expressed as number of cells per mm2. Tissue sections were counterstained with cresyl-violet and immunopositive labeling was quantitated only when associated with a cell nucleus.2. Immunocytochemistry for CCL-2Before immunohistochemistry, antigens were retrieved from Bouin’s-fixed tissue by microwaving for 11 minutes at 650 W. Ten-um paraffin wax sections were stained with anti-rat CCL-2 goat polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA) using standard methodology. In double-labeling studies, sections were initially stained for CCL-2, which was revealed using diaminobenzidine as the chromogen (brown precipitate), before using cell identification markers for ED-1 which was revealed by very intense purple (VIP) (Vector Laboratories, Peterborough, UK) (purple precipitate). Amendments were required for the identification of CCL-2 in liver. Formalin-fixed liver sections were dewaxed and rehydrated using standard procedures. Antigens for the antibodies ED-1 and HB19926 were revealed using a pressure cooker (121C for 20 minutes). Sections were incubated for 1 hour in the respective antibodies according to the manufacturers’ instructions. Labeling for cell markers was revealed using directly horseradish peroxidase-conjugated secondary antibodies to mouse or rabbit IgG (Sigma-Aldrich) to eliminate labeling of endogenous hepatic biotin. Retrieval of CCL-2 antigens in the liver required an additional pretreatment (0.04% pepsin in 0.1% HCl for 20 minutes) before overnight incubation at 4C in anti-CCL-2 rabbit polyclonal antibody (AbCam Ltd., Cambridge, UK). CCL-2 antibody was detected using biotinylated anti-rabbit IgG and standard ABC (Vector Laboratories) because amplification was necessary to reveal CCL-2. Single-labeled and double-labeled immunohistochemistry in the liver was revealed using diaminobenzidine and VIP as described above.
  Other Reagents Used: Dexamethasone 21-phosphate, EDTA
  Number Of Protocols: 2
• Reference: 1. Damoiseaux, J.G. et al. (1994) Rat macrophage lysosomal membrane antigen recognised by monoclonal antibody ED1. Immunol. 83: 140-147.
  2. Dijkstra, C.D. et al. (1985) The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in the rat recognised by monoclonal antibodies ED1, ED2 and ED3. Immunol 54: 589-599.
  3. Bauer, J. et al. (1994) Phagocytic activity of macrophages and microglial cells during the course of Acute and Chronic Relapsing Experimental Autoimmune Encephalomyelitis. J. Neurosci. Res. 38: 365-375.
  4. Bao, F. et al. (2004) Early anti-inflammatory treatment reduces lipid peroxidation and protein nitration after spinal cord injury in rats. J, Neuro-chem. 88:1335-1344.
  5. Zilka, N. et al. (2009) Human misfolded truncated tau protein promotes activation of microglia and

Target Info

• Gene Symbol: Cd68
• Alias Symbols: MGC114377, Cd68
• NCBI Gene Id: 287435
• Protein Name: Cd68 molecule EMBL AAH98931.1
• Description of Target: MOUSE ANTI RAT CD68
  
• Uniprot ID: Q4FZY1
• Protein Accession #: NP_001026808.1
• Protein Size (# AA): 330