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86791
Suppressive Myeloid Cell Phenotyping IHC Antibody Sampler Kit
Primary Antibodies

Suppressive Myeloid Cell Phenotyping IHC Antibody Sampler Kit #86791

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Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin lymphoma using CD14 (D7A2T) Rabbit mAb (IHC Formulated) performed on the Leica® Bond™ Rx.

Immunohistochemical analysis of paraffin-embedded human endometrioid carcinoma using CD11b/ITGAM (D6X1N) Rabbit mAb performed on the Leica® Bond™ Rx.

Flow cytometric analysis of Jurkat cells (blue) and THP-1 cells (green) using CD68 (D4B9C) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Immunohistochemical analysis of paraffin-embedded human tonsil using CD68 (D4B9C) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using CD163 (D6U1J) Rabbit mAb performed on the Leica® Bond™ Rx.

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using CD206/MRC1 (E2L9N) Rabbit mAb performed on the Leica® BOND™ Rx.

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using M-CSF Receptor (E4T8Z) Rabbit mAb performed on the Leica® BOND™ Rx.

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Arginase-1 (D4E3M™) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

Flow cytometric analysis of human whole blood using Arginase-1 (D4E3M™) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Analysis was performed on cells in the granulocyte gate.

Flow cytometric analysis of Jurkat cells (blue) and Raji cells (green) using MHC Class II (LGII-612.14) Mouse mAb (solid lines) or a concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.

Flow cytometric analysis of unpermeabilized NIH/3T3 cells (blue) and unpermeabilized F9 cells (green) using CD15/SSEA1 (MC480) Mouse mAb.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CD14 (D7A2T) Rabbit mAb (IHC Formulated).

Immunohistochemical analysis of paraffin-embedded human clear cell adenocarcinoma of the ovary using CD11b/ITGAM (D6X1N) Rabbit mAb.

Flow cytometric analysis of live human peripheral blood mononuclear cells co-stained with anti-human CD14 using CD68 (D4B9C) XP® Rabbit mAb (right) compared to a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using CD68 (D4B9C) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human clear cell adenocarcinoma of the ovary using CD163 (D6U1J) Rabbit mAb performed on the Leica® Bond™ Rx.

Immunohistochemical analysis of paraffin-embedded MUTZ-3 cell pellet (left, positive) or THP-1 cell pellet (right, negative) using CD206/MRC1 (E2L9N) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using M-CSF Receptor (E4T8Z) Rabbit mAb.

Confocal immunofluorescent analysis of mouse liver (positive; left) or small intestine (negative; right) using Arginase-1 (D4E3M™) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5 #4084 (fluorescent DNA dye).

Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using MHC Class II (LGII-612.14) Mouse mAb performed on the Leica® BOND™ Rx.

Confocal immunofluorescent analysis of mouse embryonic stem cells growing on mouse embryonic fibroblast (MEF) feeder cells using CD15/SSEA1 (MC480) Mouse mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using CD14 (D7A2T) Rabbit mAb (IHC Formulated).

Immunohistochemical analysis of paraffin-embedded THP-1 cell pellet (left, positive) or Jurkat cell pellet (right, negative) using CD11b/ITGAM (D6X1N) Rabbit mAb.

Flow cytometric analysis of fixed and permeabilized human peripheral blood mononuclear cells co-stained with anti-human CD14 using CD68 (D4B9C) XP® Rabbit mAb (right) compared to a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using CD68 (D4B9C) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CD163 (D6U1J) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CD206/MRC1 (E2L9N) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using M-CSF Receptor (E4T8Z) Rabbit mAb (left) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).

Confocal immunofluorescent analysis of mouse primary bone marrow-derived macrophages (BMDMs) using Arginase-1 (D4E3M™) XP® Rabbit mAb (green). BMDMs were differentiated with M-CSF (20 ng/ml, 7 days) and activated with either IL-4/cAMP (20 ng/ml, 0.5 mM, 24 hours; left) or LPS/IFNγ (50 ng/ml, 20 ng/ml, 24 hours; right). Red = Propidium Iodide (PI)/RNase Staining Solution #4087.

Immunohistochemical analysis of paraffin-embedded human gastric carcinoma using MHC Class II (LGII-612.14) Mouse mAb performed on the Leica® BOND™ Rx.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CD15/SSEA1 (MC480) Mouse mAb.

Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using CD14 (D7A2T) Rabbit mAb (IHC Formulated).

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CD11b/ITGAM (D6X1N) Rabbit mAb.

Confocal immunofluorescent analysis of THP-1 (left) and Jurkat (right) cells using CD68 (D4B9C) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Projected images from a z-stack series are shown.

Immunohistochemical analysis of paraffin-embedded THP-1 (left) and Jurkat (right) cell pellets using CD68 (D4B9C) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using CD163 (D6U1J) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using CD206/MRC1 (E2L9N) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded GDM-1 cell pellet (left, positive) or HeLa cell pellet (right, negative) using M-CSF Receptor (E4T8Z) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using MHC Class II (LGII-612.14) Mouse mAb.

Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using CD15/SSEA1 (MC480) Mouse mAb.

Immunohistochemical analysis of paraffin-embedded THP-1 cell pellet (left, positive) or Raji cell pellet (right, negative) using CD14 (D7A2T) Rabbit mAb (IHC Formulated).

Western blot analysis of extracts from various cells using CD11b/ITGAM (D6X1N) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Multiplex immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using CD68 (D4B9C) XP® rabbit mAb (red), PD-1 (EH33) mouse mAb #43248 (green), CD8α (C8/144B) mouse mAb #70306 (magenta), Pan-keratin (C11) mouse mAb #4545 (cyan), LAG3 (D2G4O™) XP® rabbit mAb #15372 (orange), and TIM-3 (D5D5R™) XP® rabbit mAb #45208 (yellow).

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using CD163 (D6U1J) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using CD206/MRC1 (E2L9N) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using M-CSF Receptor (E4T8Z) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Arginase-1 (D4E3M™) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using MHC Class II (LGII-612.14) Mouse mAb (left) compared to concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (right).

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using CD15/SSEA1 (MC480) Mouse mAb.

Multiplex immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using CD68 (D4B9C) XP® rabbit mAb (orange), PD-L1 (E1L3N®) XP® rabbit mAb #13684 (red), PD-L2 (D7U8C™) XP® rabbit mAb (magenta) #82723, Arginase-1 (D4E3M™) XP® rabbit mAb #93668 (green), IDO (D5J4E™) rabbit mAb #86630 (yellow), and Pan-keratin (C11) mouse mAb #4545 (cyan).

Immunohistochemical analysis of paraffin-embedded human serous carcinoma of the ovary using CD163 (D6U1J) Rabbit mAb.

Western blot analysis of extracts from various cell lines using CD206/MRC1 (E2L9N) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western blot analysis of extracts from various human cells using M-CSF Receptor (E4T8Z) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Immunohistochemical analysis of paraffin-embedded normal human liver using Arginase-1 (D4E3M™) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using MHC Class II (LGII-612.14) Mouse mAb.

IHC analysis of paraffin-embedded mouse embryo, showing staining of hair follicles, using CD15/SSEA1 (MC480) Mouse mAb.

Immunohistochemical analysis of paraffin-embedded SU-DHL-1 cell pellet (left, positive) or A-375 cell pellet (right, negative) using CD163 (D6U1J) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Arginase-1 (D4E3M™) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using MHC Class II (LGII-612.14) Mouse mAb.

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using CD15/SSEA1 (MC480) Mouse mAb.

Western blot analysis of extracts from SU-DHL-1 and A-375 cells using CD163 (D6U1J) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower)

Immunohistochemical analysis of paraffin-embedded mouse liver using Arginase-1 (D4E3M™) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using MHC Class II (LGII-612.14) Mouse mAb.

Western blot analysis of extracts from mouse and rat liver using Arginase-1 (D4E3M™) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded Raji cell pellet (left, positive) or Jurkat cell pellet (right, negative) using MHC Class II (LGII-612.14) Mouse mAb.

Western blot analysis of extracts from mouse liver and mouse small intestine using Arginase-1 (D4E3M™) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Immunoprecipitation of MHC Class II from Raji cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is MHC Class II (LGII-612.14) Mouse mAb. Western blot analysis was performed using MHC Class II (LGII-612.14) Mouse mAb. Anti-mouse IgG, HRP-linked Antibody #7076 was used as the secondary antibody.

Western blot analysis of extracts from various cell lines using MHC Class II (LGII-612.14) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

To Purchase # 86791T
Product # Size Price
86791T
1 Kit  (9 x 20 µl) N/A

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
CD14 (D7A2T) Rabbit mAb (IHC Formulated) 75181 20 µl
  • IHC
H Rabbit IgG
CD11b/ITGAM (D6X1N) Rabbit mAb 49420 20 µl
  • WB
  • IHC
H 170 Rabbit IgG
CD68 (D4B9C) XP® Rabbit mAb 76437 20 µl
  • IHC
  • IF
  • F
H Rabbit IgG
CD163 (D6U1J) Rabbit mAb 93498 20 µl
  • WB
  • IHC
H 160, 170 Rabbit IgG
CD206/MRC1 (E2L9N) Rabbit mAb 91992 20 µl
  • WB
  • IHC
H 190-250 Rabbit IgG
M-CSF Receptor (E4T8Z) Rabbit mAb 28917 20 µl
  • WB
  • IHC
H 140-200 Rabbit IgG
Arginase-1 (D4E3M™) XP® Rabbit mAb 93668 20 µl
  • WB
  • IHC
  • IF
  • F
H M R 40 Rabbit IgG
MHC Class II (LGII-612.14) Mouse mAb 68258 20 µl
  • WB
  • IP
  • IHC
  • F
H 25-35, 50-65 Mouse IgG1
CD15/SSEA1 (MC480) Mouse mAb 4744 20 µl
  • IHC
  • IF
  • F
H M N/A Mouse IgM

Product Description

The Suppressive Myeloid Cell Phenotyping IHC Antibody Sampler Kit provides an economical means of detecting the accumulation of immune cell types in formalin-fixed, paraffin-embedded tissue samples.

Specificity / Sensitivity

Each antibody included in the Suppressive Myeloid Cell Phenotyping IHC Antibody Sampler Kit recognizes endogenous levels of its target human protein. M-CSF Receptor (E4T8Z) Rabbit mAb cross-reacts with an unidentified protein of 70 kDa in some cell extracts. Arginase-1 (D4E3M™) XP® Rabbit mAb does not cross-react with arginase-2. MHC Class II (LGII-612.14) Mouse mAb exhibits strong reactivity with HLA-DRB and weak reactivity with HLA-DPB in cell lines transfected with constructs expressing Myc/DDK-tagged HLA-DRB and HLA-DPB, respectively. Reactivity is not observed with HLA-DMB, HLA-DOB, and HLA-DQB in cell lines transfected with constructs expressing Myc/DDK-tagged HLA-DMB, HLA-DOB, and HLA-DQB.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro319 of human CD14 protein or Val47 of human arginase-1 protein, or with recombinant protein specific to human CD68 protein, human CD163 protein, human CD206/MRC1 protein, the carboxy terminus of human M-CSF receptor protein, or the amino terminus of human CD11b/ITGAM protein. CD15/SSEA1 (MC480) Mouse mAb is produced by immunizing animals with F9 teratoma cells (X-irradiated). MHC Class II (LGII-612.14) Mouse mAb is produced by immunizing animals with cultured human B lymphoid cells treated with IFN-gamma.

Background

A combination of multiple biomarkers are required to characterize the phenotype of myeloid cell lineages. Cluster of differentiation molecule 14 (CD14) is a leucine-rich repeat-containing pattern recognition receptor with expression largely restricted to the monocyte/macrophage cell lineage (1), but can be unregulated on polymorphonuclear as well as nonmyeloid cells such as B cells and gingival fibroblasts (2,3). CD11b (Integrin alpha M or ITGAM) is a transmembrane protein forming heterodimers that are composed of α and β subunits (4). CD11b is expressed by, and commonly used as a marker for myeloid lineage cells, including neutrophils, monocytes, macrophages, dendritic cells, and microglia (5), but has also been detected on a subset of B cells (6-8). CD68 (macrosialin) is a heavily glycosylated transmembrane protein that is expressed by and commonly used as a marker for monocytes and macrophages (9,10), but there is also evidence of non-myeloid cell expression (11). The CD15 carbohydrate epitope is preferentially expressed in mature human neutrophils, monocytes, and all myeloid cells from the promyelocyte stage onwards, making it a useful cell surface marker (12-14). It is also expressed in some tissues, such as epithelial cells of intestinal tissues (15,16), and in certain neurons and glial cells in the central nervous system (17).  

CD163 is a transmembrane scavenger receptor expressed on the macrophage surface. It has 9 B-type SRCR extracellular domains mediating serum haptoglobin clearing/endocytosis, pathogen binding and signal transduction, and calcium binding (18,19). The mannose receptor (CD206/MR/CLEC13D/MMR/MRC1/Macrophage mannose receptor 1) is an endocytic receptor expressed by populations of dendritic cells, macrophages, and nonvascular endothelium (20). CD206/MRC1 receptor functions include a role in antigen cross-presentation, clearance of endogenous proteins, pathogen detection and trafficking through lymphatic vessels (21-24). Macrophage-colony stimulating factor (M-CSF, CSF-1) receptor is an integral membrane tyrosine kinase encoded by the c-fms proto-oncogene. M-CSF receptor is expressed in monocytes (macrophages and their progenitors) and drives growth and development of this blood cell lineage (25,26). CD163, CD206, and M-CSF receptors are used as surface markers of M2 type macrophages, including M2 type tumor associated macrophages (TAMs), which facilitate cancer progression by secreting cytokines to promote angiogenesis, immunosuppression, and metastasis (20,27,28). Arginase-1 catalyzes the final step of the urea cycle converting L-arginine to L-ornithine and urea (29). Myeloid-derived suppressor cells express high levels of arginase-1, increasing the catabolism of L-arginine resulting in L-arginine depletion in the inflammatory microenvironment of cancer. The reduced availability of L-arginine suppresses T cell proliferation and function and thus contributes to tumor progression (30,31).

Major histocompatibility complex class II (MHC class II) molecules are heterodimeric, transmembrane glycoproteins expressed on the surface of antigen-presenting cells such as macrophages, dendritic cells, and B cells. Expression can also be induced through interferon-γ signaling (32). Prior to being displayed on the cell membrane, MHC class II molecules are loaded with exogenous peptide antigens approximately 15-24 amino acids in length that were derived from endocytosed extracellular proteins digested in the lysosome. Antigen-presentation through MHC class II is required for T cell activation during the immune response to extracellular pathogens (33). High expression of MHC class II on myeloid cell lineages is used as a surface marker of M1 type macrophages, including M1 type TAMs, which can assist in tumor eradication by secreting cytokines to activate anti-tumor immune responses, and inhibit angiogenesis and metastasis (27,28).

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  18. Graversen, J.H. and Moestrup, S.K. (2015) Membranes (Basel) 5, 228-52.
  19. Etzerodt, A. and Moestrup, S.K. (2013) Antioxid Redox Signal 18, 2352-63.
  20. Martinez-Pomares, L. (2012) J Leukoc Biol 92, 1177-86.
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  26. Bourette, R.P. and Rohrschneider, L.R. (2000) Growth Factors 17, 155-66.
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  28. Mills, C.D. et al. (2000) J Immunol 164, 6166-73.
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  30. Gabrilovich, D.I. and Nagaraj, S. (2009) Nat Rev Immunol 9, 162-74.
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Pathways & Proteins

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For Research Use Only. Not For Use In Diagnostic Procedures.

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