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PRODUCT CODE: EM1901-94

CD68 Mouse Monoclonal Antibody [A3C3] (EM1901-94)

  • IVD–IHC

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

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Western blot analysis of CD68 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-94, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br /> Positive control: <br /> Lane 1: A431 cell lysate<br /> Lane 2: THP-1 cell lysate
  • Western blot analysis of CD68 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-94, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br /> Positive control: <br /> Lane 1: A431 cell lysate<br /> Lane 2: THP-1 cell lysate
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD68 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-94, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CD68 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-94, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue using anti-CD68 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-94, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of CD68 was done on THP-1 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-94, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of CD68 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-94, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: A431 cell lysate
Lane 2: THP-1 cell lysate

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

CD68 Mouse Monoclonal Antibody [A3C3] (EM1901-94)

Immunogen

Synthetic peptide within human cd68 aa 320-354.

Host

Mouse

Positive Control

A431 cell lysate, THP-1 cell lysate, human tonsil tissue, human spleen tissue, human lung cancer tissue, THP-1.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

A3C3

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein G affinity purified.

MOLECULAR WEIGHT

Predicted band size: 37 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB:1:500-1:1,000

  • IHC-P:1:50-1:200

  • FC:1:50-1:100

TARGET

UNIPROT #

P34810

PROTEIN NAME

CD68

SYNONYMS

CD 68 antibody; CD68 antibody; CD68 antigen antibody; CD68 molecule antibody; CD68_HUMAN antibody; DKFZp686M18236 antibody; gp11 antibody; Gp110 antibody; LAMP4 antibody; Macrophage antigen CD68 (microsialin) antibody; MACROPHAGE ANTIGEN CD68 antibody; Macrosialin antibody; SCARD1 antibody; Scavenger receptor class D member 1 antibody

SEQUENCE SIMILARITIES

Belongs to the LAMP family.

TISSUE SPECIFICITY

Highly expressed by blood monocytes and tissue macrophages. Also expressed in lymphocytes, fibroblasts and endothelial cells. Expressed in many tumor cell lines which could allow them to attach to selectins on vascular endothelium, facilitating their dissemination to secondary sites.

POST-TRANSLATIONAL MODIFICATION

N- and O-glycosylated.

SUBCELLULAR LOCATION

Cell membrane. Endosome membrane, lysosome membrane.

FUNCTION

CD68 (Cluster of Differentiation 68) is a protein highly expressed by cells in the monocyte lineage (e.g., monocytic phagocytes, osteoclasts), by circulating macrophages, and by tissue macrophages (e.g., Kupffer cells, microglia). Human CD68 is a transmembrane glycoprotein, heavily glycosylated in its extracellular domain, with a molecular weight of 110 kD. Its primary sequence consists of 354 amino acids with predicted molecular weight of 37.4 kD if it were not glycosylated. Immunohistochemistry can be used to identify the presence of CD68, which is found in the cytoplasmic granules of a range of different blood cells and myocytes. It is particularly useful as a marker for the various cells of the macrophage lineage, including monocytes, histiocytes, giant cells, Kupffer cells, and osteoclasts. This allows it to be used to distinguish diseases of otherwise similar appearance, such as the monocyte/macrophage and lymphoid forms of leukaemia (the latter being CD68 negative). Its presence in macrophages also makes it useful in diagnosing conditions related to proliferation or abnormality of these cells, such as malignant histiocytosis, histiocytic lymphoma, and Gaucher's disease. Anti-CD68 monoclonal antibodies that react with tissues of rodent and other species include ED1, FA-11, KP1 (a.k.a. C68/684), 6A326, 6F3, 12E2, 10B1909, and SPM130. Monoclonals that react with humans include, Ki-M7, PG-M1, 514H12, ABM53F5, 3F7C6, 3F7D3, Y1/82A, EPR20545, CDLA68-1, LAMP4-824.