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37495
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit #37495

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Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 1
Immunofluorescent analysis of mouse spleen using FoxP3 (D6O8R) Rabbit mAb (red) and a CD4 antibody (green). Blue = DRAQ5® #4084 (fluorescent DNA dye).

Note: FoxP3 (D6O8R) Rabbit mAb has been validated on freshly-prepared frozen tissue sections that were fixed for 15 minutes in 4% formaldehyde. Over-fixation diminishes the performance of this antibody.

Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 2
Immunohistochemical analysis of paraffin-embedded 4T1 metastatic tumor in mouse lung using FoxP3 (D6O8R) Rabbit mAb performed on the Leica® BOND™ Rx.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 3
Immunohistochemical analysis of paraffin-embedded mouse thymus using FoxP3 (D6O8R) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 4
Immunohistochemical analysis of paraffin-embedded mouse spleen using FoxP3 (D6O8R) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 5
Immunohistochemical analysis of paraffin-embedded mouse small intestine using FoxP3 (D6O8R) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 6
Immunohistochemical analysis of paraffin-embedded mouse lung using FoxP3 (D6O8R) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 7
Flow cytometric analysis of mouse splenocytes using FoxP3 (D6O8R) Rabbit mAb (right) and co-stained with CD4 (RM4-5) Rat mAb (APC Conjugate) #82116, 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) was used as a secondary antibody.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 8
Immunohistochemical analysis of paraffin-embedded 4T1 metastatic tumor in mouse lung using CD4 (D7D2Z) Rabbit mAb performed on the Leica® BOND™ Rx.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 9
Western blot analysis of extracts from various cell lines and tissues using CD4 (D7D2Z) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 10
Immunohistochemical analysis of paraffin-embedded mouse colon using CD4 (D7D2Z) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 11
Immunohistochemical analysis of paraffin-embedded mouse small intestine using CD4 (D7D2Z) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 12
Immunohistochemical analysis of paraffin-embedded mouse spleen using CD4 (D7D2Z) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 13
Immunohistochemical analysis of paraffin-embedded rat lung using CD4 (D7D2Z) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 14
Immunohistochemical analysis of paraffin-embedded human colon adenocarinoma using Granzyme B (D6E9W) Rabbit mAb performed on the Leica® BOND™ Rx.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 15
Immunohistochemical analysis of paraffin-embedded human papillary carcinoma of the breast using Granzyme B (D6E9W) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 16
Immunohistochemical analysis of paraffin-embedded KARPAS-299 cell pellet (left, positive) or Jurkat cell pellet (right, negative) using Granzyme B (D6E9W) Rabbit mAb. KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 17
Western blot analysis of extracts from various human cells using Granzyme B (D6E9W) Rabbit mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower). CD8+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of human interleukin-2 (hIL-2) #8907 (20 ng/ml). KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 18
Western blot analysis of extracts from CTLL-2, mouse CD8+ T, and C2C12 cells using Granzyme B (D6E9W) Rabbit mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower). CD8+ T cells were purified from mouse spleens and stimulated for 9 days using beads coated with mouse CD3 and CD28 antibodies in the presence of Mouse Interleukin-2 (mIL-2) #5201 (20 ng/ml).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 19
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Granzyme B (D6E9W) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using Granzyme B (D6E9W) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 21
Immunohistochemical analysis of paraffin-embedded mouse liver using F4/80 (D2S9R) XP® Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 22
Western blot analysis of extracts from various cell lines using F4/80 (D2S9R) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 23
Immunohistochemical analysis of paraffin-embedded mouse spleen using F4/80 (D2S9R) XP® Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 24
Immunohistochemical analysis of paraffin-embedded mouse small intestine using F4/80 (D2S9R) XP® Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 25
Immunohistochemical analysis of paraffin-embedded Raw 264.7 cell pellet (left, positive) or Neuro-2A cell pellet (right, negative) using F4/80 (D2S9R) XP® Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 26
Multiplex immunohistochemical analysis of paraffin-embedded mouse LL2 syngeneic tumor tissue using F4/80 (D2S9R) XP® rabbit mAb (orange), PD-1 (D7D5W) XP® rabbit mAb #84651 (green), PD-L1 (D5V3B) rabbit mAb #64988 (yellow), CD3ε (D4V8L) rabbit mAb #99940 (red), CD8α (D4W2Z) XP® rabbit mAb #98941 (magenta) and pan keratin (C11) mouse mAb #4279 (cyan).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 27
Immunohistochemical analysis of paraffin-embedded human endometrioid carcinoma using CD19 (Intracellular Domain) (D4V4B) XP® Rabbit mAb performed on the Leica® Bond™ Rx.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 28
Western blot analysis of extracts from various cell lines using CD19 (Intracellular Domain) (D4V4B) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, CD19 protein is not detected in either U-937 cells or Raw 264.7 cells.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 29
Western blot analysis of extracts from Daudi cells, untreated (-) or treated with PNGase F (+), using CD19 (Intracellular Domain) (D4V4B) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 30
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CD19 (Intracellular Domain) (D4V4B) XP® Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 31
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using CD19 (Intracellular Domain) (D4V4B) XP® Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded human lymphoma using CD19 (Intraceullular Domain) (D4V4B) XP® Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 33
Immunohistochemical analysis of paraffin-embedded mouse spleen using CD19 (Intracellular Domain) (D4V4B) XP® Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 34
Immunohistochemical analysis of paraffin-embedded human tonsil using CD19 (Intracellular Domain) (D4V4B) XP® Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 35
Immunohistochemical analysis of paraffin-embedded 4T1 mammary tumor using CD11c (D1V9Y) Rabbit mAb performed on the Leica® Bond™ Rx.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 36
Western blot analysis of extracts from Raw 264.7, C2C12, and 3T3 cells using CD11c (D1V9Y) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 37
Immunohistochemical analysis of paraffin-embedded mouse kidney using CD11c (D1V9Y) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 38
Immunohistochemical analysis of paraffin-embedded mouse liver using CD11c (D1V9Y) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 39
Immunohistochemical analysis of paraffin-embedded mouse lung using CD11c (D1V9Y) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 40
Immunohistochemical analysis of paraffin-embedded Raw 264.7 cell pellet (left, positive) or C2C12 cell pellet (right, negative) using CD11c (D1V9Y) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 41
Immunohistochemical analysis of paraffin-embedded mouse spleen using CD11c (D1V9Y) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 42
Confocal immunofluorescent analysis of mouse spleen (left) and mouse kidney (right) using CD11c (D1V9Y) Rabbit mAb. Nuclei were labeled with ProLong® Gold Antifade reagent with DAPI #8961 (blue).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 43
Confocal immunofluorescent analysis of Raw 264.7 cells (left, positive) and 3T3 cells (right, negative) using CD11c (D1V9Y) Rabbit mAb (green). Red = Propidium Iodide (PI)/RNase Staining Solution #4087.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 44
Immunohistochemical analysis of paraffin-embedded mouse lung using CD8α (D4W2Z) XP® Rabbit mAb (Mouse Specific) performed on the Leica® BOND™ Rx.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 45
Western blot analysis of extracts from mouse CD8+ T cells, mouse spleen, EL4 cells, and Raw264.7 cells using CD8α (D4W2Z) XP® Rabbit mAb (Mouse Specific) (upper), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 46
Immunohistochemical analysis of paraffin-embedded mouse spleen using CD8α (D4W2Z) XP® Rabbit mAb (Mouse Specific).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 47
Immunohistochemical analysis of paraffin-embedded mouse small intestine using CD8α (D4W2Z) XP® Rabbit mAb (Mouse Specific).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 48
Multiplex immunohistochemical analysis of paraffin-embedded mouse LL2 syngeneic tumor tissue using CD8α (D4W2Z) XP® rabbit mAb (magenta), F4/80 (D2S9R) XP® rabbit mAb #70076 (orange), PD-1 (D7D5W) XP® rabbit mAb #84651 (green), PD-L1 (D5V3B) rabbit mAb #64988 (yellow), CD3ε (D4V8L) rabbit mAb #99940 (red) and pan keratin (C11) mouse mAb #4279 (cyan).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 49
Immunohistochemical analysis of paraffin-embedded mouse small intestine using CD3ε (D4V8L) Rabbit mAb performed on the Leica® BOND™ Rx.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 50
Western blot analysis of extracts from mouse spleen, EL4 cells, and C2C12 cells using CD3ε (D4V8L) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 51
Immunohistochemical analysis of paraffin-embedded mouse spleen using CD3ε (D4V8L) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 52
Immunohistochemical analysis of paraffin-embedded mouse small intestine using CD3ε (D4V8L) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 53
Immunohistochemical analysis of paraffin-embedded mouse lung using CD3ε (D4V8L) Rabbit mAb.
Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit: Image 54
Multiplex immunohistochemical analysis of paraffin-embedded mouse LL2 syngeneic tumor tissue using CD3ε (D4V8L) Rabbit mAb (red), F4/80 (D2S9R) XP® Rabbit mAb #70076 (orange), PD-1 (D7D5W) XP® Rabbit mAb #84651 (green), PD-L1 (D5V3B) Rabbit mAb #64988 (yellow), CD8α (D4W2Z) XP® Rabbit mAb #98941 (magenta) and Pan-Keratin (C11) Mouse mAb (Biotinylated) #4279 (cyan). Note: Biotinylated Pan-Keratin (C11) Mouse mAb was used in order to avoid mouse on mouse background staining.
To Purchase # 37495T
Product # Size Price
37495T
1 Kit  (8 x 20 microliters) $ 658

Product Description

The Mouse Immune 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 Mouse Immune Cell Phenotyping IHC Antibody Sampler Kit recognizes endogenous levels of its target mouse protein. Non-specific staining has been observed in mouse pancreas with CD3ε (D4V8L) Rabbit mAb, in mouse kidney and liver with CD4 (D7D2Z) Rabbit mAb, and in the sweat glands with Granzyme B (D6E9W) Rabbit mAb.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Val31 of mouse CD3ε protein, Asp42 of mouse CD8α protein, Ala232 of mouse CD4 protein, Pro44 of mouse FoxP3 protein, Leu427 of human CD19 protein, Ala1153 of mouse CD11c protein, or with recombinant proteins specific to human Granzyme B protein or mouse F4/80 protein.

Background

Cluster of Differentiation 3 (CD3) is a multiunit protein complex expressed on the surface of T-cells that directly associates with the T-cell receptor (TCR). CD3 is composed of four polypeptides: ζ, γ, ε and δ. Engagement of TCR complex with antigens presented in Major Histocompatibility Complexes (MHC) induces tyrosine phosphorylation in the immunoreceptor tyrosine-based activation motif (ITAM) of CD3 proteins. CD3 phosphorylation is required for downstream signaling through ZAP-70 and p85 subunit of PI-3 kinase, leading to T cell activation, proliferation, and effector functions (1). Cluster of Differentiation 8 (CD8) is a transmembrane glycoprotein expressed primarily on cytotoxic T cells, but has also been described on a subset of dendritic cells in mice (2,3). On T cells, CD8 is a co-receptor for the TCR, and these two distinct structures are required to recognize antigen bound to MHC Class I (2). Cluster of Differentiation 4 (CD4) is expressed on the surface of T helper cells, regulatory T cells, monocytes, macrophages, and dendritic cells, and plays an important role in the development and activation of T cells. On T cells, CD4 is the co-receptor for the TCR, and these two distinct structures recognize antigen bound to MHC Class II. CD8 and CD4 co-receptors ensure specificity of the TCR–antigen interaction, prolong the contact between the T cell and the antigen presenting cell, and recruit the tyrosine kinase Lck, which is essential for T cell activation (2). Granzyme B is a serine protease expressed by CD8+ cytotoxic T lymphocytes and natural killer (NK) cells and is a key component of the immune response to pathogens and transformed cancer cells (4). Forkhead box P3 (FoxP3) is crucial for the development of T cells with immunosuppressive regulatory properties and is a well-established marker for T regulatory cells (Tregs) (5). CD19 is a co-receptor expressed on B cells that amplifies the signaling cascade initiated by the B cell receptor (BCR) to induce activation. It is a biomarker of B lymphocyte development, lymphoma diagnosis, and can be utilized as a target for leukemia immunotherapies (6,7). F4/80 (EMR1) is a heavily glycosylated G-protein-coupled receptor and is a well-established marker for mouse macrophages (8). CD11c (integrin αX, ITGAX) is a transmembrane glycoprotein highly expressed by dendritic cells, and has also been observed on activated NK cells, subsets of B and T cells, monocytes, granulocytes, and some B cell malignancies including hairy cell leukemia (9,10).
  1. Kuhns, M.S. et al. (2006) Immunity 24, 133-9.
  2. Zamoyska, R. (1994) Immunity 1, 243-6.
  3. Shortman, K. and Heath, W.R. (2010) Immunol Rev 234, 18-31.
  4. Trapani, J.A. (2001) Genome Biol 2, REVIEWS3014.
  5. Ochs, H.D. et al. (2007) Immunol Res 38, 112-21.
  6. Tedder, T.F. et al. (1997) Immunity 6, 107-18.
  7. Scheuermann, R.H. and Racila, E. (1995) Leuk Lymphoma 18, 385-97.
  8. McKnight, A.J. et al. (1996) J Biol Chem 271, 486-9.
  9. Kohrgruber, N. et al. (1999) J Immunol 163, 3250-9.
  10. Qualai, J. et al. (2016) PLoS One 11, e0154253.

Pathways & Proteins

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