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49938
Microglia Proliferation Module Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Microglia Proliferation Module Antibody Sampler Kit #49938

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Microglia Proliferation Module Antibody Sampler Kit: Image 1

Flow cytometric analysis of NIH/3T3 cells (red) and 32D cells (blue), using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 2

Flow cytometric analysis of Raw264.7 cells (blue) and J774A.1 cells (green) using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Microglia Proliferation Module Antibody Sampler Kit: Image 3

Flow cytometric analysis of Jurkat cells using Ki-67 (D3B5) Rabbit mAb and Propidium Iodide (PI)/RNase Staining Solution #4087. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Microglia Proliferation Module Antibody Sampler Kit: Image 4

Chromatin immunoprecipitations were performed with cross-linked chromatin from HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes, and either MCM2 (D7G11) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human TAP1 Promoter Primers #5148, human IRF-1 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Microglia Proliferation Module Antibody Sampler Kit: Image 5

Flow cytometric analysis of untreated Jurkat cells using Survivin (71G4B7E) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

Microglia Proliferation Module Antibody Sampler Kit: Image 6

Confocal immunofluorescent analysis of HeLa cells using HP1α (C7F11) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

Microglia Proliferation Module Antibody Sampler Kit: Image 7

Flow cytometric analysis of untreated Jurkat cells using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) XP® Rabbit mAb compared to propidium iodide (DNA content). The boxed population indicates phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198)-positive cells.

Microglia Proliferation Module Antibody Sampler Kit: Image 8

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Microglia Proliferation Module Antibody Sampler Kit: Image 9

Confocal immunofluorescent analysis of mouse Tg2576 brain which overexpresses mutant human APP695. Sections were first labeled with HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) (green) and APP/β-Amyloid (NAB228) Mouse mAb #2450 (yellow). After blocking free secondary binding sites with Mouse (G3A1) mAb IgG1 Isotype Control #5415, sections were incubated with GFAP (GA5) Mouse mAb (Alexa Fluor® 647 Conjugate) #3657 (red). Nuclei were labeled with Hoechst 33342 #4082 (blue).

Microglia Proliferation Module Antibody Sampler Kit: Image 10

Confocal immunofluorescent analysis of mouse Tg2576 brain which overexpresses mutant human APP695. Sections were first labeled with ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) #67824 (green) and APP/β-Amyloid (NAB228) Mouse mAb #2450 (yellow). After blocking free secondary binding sites with Mouse (G3A1) mAb IgG1 Isotype Control #5415, sections were incubated with GFAP (GA5) Mouse mAb (Alexa Fluor® 647 Conjugate) #3657 (red). Nuclei were labeled with Hoechst 33342 #4082 (blue).

Microglia Proliferation Module Antibody Sampler Kit: Image 11

Confocal immunofluorescent analysis of the ventricular zone in P21 mouse brain using Ki-67 (D3B5) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Microglia Proliferation Module Antibody Sampler Kit: Image 12

Confocal immunofluorescent analysis of HeLa cells using MCM2 (D7G11) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Microglia Proliferation Module Antibody Sampler Kit: Image 13

Confocal immunofluorescent analysis of HeLa cells using Survivin (71G4B7E) Rabbit mAb (green). Mitochondria have been labeled with MitoTracker® Red CMXRos (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Microglia Proliferation Module Antibody Sampler Kit: Image 14

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear localization using HP1 α (C7F11) Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 15

Confocal immunofluorescent analysis of HT-1080 cells using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) XP® Rabbit mAb (green), β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red), and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (blue).

Microglia Proliferation Module Antibody Sampler Kit: Image 16

Confocal immunofluorescent analysis of 32D cells (left) and C2C12 cells (right), using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Microglia Proliferation Module Antibody Sampler Kit: Image 17

Confocal immunofluorescent analysis of mouse primary bone marrow-derived macrophages (BMDMs) either untreated (upper left) or treated with LPS (50 ng/ml, 4 hr, middle) or LPS followed by ATP (5 mM, 45 min, upper right), and J774A.1 (lower left) or Raw 264.7 (lower right) cells, using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Note the translocation of ASC to inflammasomes following stimulation with LPS and ATP (white arrows).

Microglia Proliferation Module Antibody Sampler Kit: Image 18

Confocal immunofluorescent analysis of HeLa cells using Ki-67 (D3B5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Microglia Proliferation Module Antibody Sampler Kit: Image 19

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using MCM2 (D7G11) XP® Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 20

Immunohistochemical analysis of frozen H1650 xenograft showing nuclear localization using Survivin (71G4B7E) Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 21

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using HP1 α (C7F11) Rabbit mAb in the presence of control peptide (left) or HP1 alpha blocking peptide #1004 (right).

Microglia Proliferation Module Antibody Sampler Kit: Image 22

Western blot analysis of extracts from HeLa, L929 and C6 cells, treated with 4 mM hydroxyurea for 20 hours to induce G1/S phase or treated with 100 nM paclitaxel or 100 ng/ml nocodazole for 20 hours to induce G2/M phase, using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D11A13) XP® Rabbit mAb (upper) or Aurora B/AIM1 Antibody #3094 (lower).

Microglia Proliferation Module Antibody Sampler Kit: Image 23

Immunohistochemical analysis of paraffin-embedded mouse spleen using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 24

Immunohistochemical analysis of paraffin-embedded J774A.1 cell pellet (left, positive) or RAW 264.7 cell pellet (right, negative) using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 25

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using MCM2 (D7G11) XP® Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 26

Immunohistochemical analysis of paraffin-embedded human transitional epithelial carcinoma of the bladder using Survivin (71G4B7E) Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 27

Western blot analysis of lysates from HeLa, NIH/3T3, C6 and COS cells using HP1 α (C7F11) Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 28

Immunohistochemical analysis of paraffin-embedded LL2 syngeneic tumor using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 29

Immunohistochemical analysis of paraffin-embedded mouse forestomach using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 30

Immunohistochemical analysis of paraffin-embedded human small intestine using MCM2 (D7G11) XP® Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 31

Immunohistochemical analysis of paraffin-embedded human lung carcinoma showing nuclear localization using Survivin (71G4B7E) Rabbit mAb #2808.

Microglia Proliferation Module Antibody Sampler Kit: Image 32

Western blot analysis of cell extracts from Baf3, 32D, and mouse spleen using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 33

Immunohistochemical analysis of paraffin-embedded mouse brain using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 34

Immunoprecipitation of MCM2 from HeLa cell lysates using MCM2 (D7G11) XP® Rabbit mAb followed by western blot using the same antibody. Lane 1 is 5% input.

Microglia Proliferation Module Antibody Sampler Kit: Image 35

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Survivin (71G4B7E) Rabbit mAb in the presence of control peptide (left) or Survivin Blocking Peptide #1037 (right).

Microglia Proliferation Module Antibody Sampler Kit: Image 36

Immunohistochemical analysis of paraffin-embedded mouse colon using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).

Microglia Proliferation Module Antibody Sampler Kit: Image 37

Western blot analysis of extracts from various cell types using MCM2 (D7G11) XP® Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 38

Immunohistochemical analysis of paraffin-embedded human pituitary adenoma using Survivin (71G4B7E) Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 39

Immunohistochemical analysis of paraffin-embedded mouse thymus using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 40

Western blot analysis of lysates from Raji (human), BaF3 (mouse) and C6 (rat) cell lines using Survivin (71G4B7E) Rabbit mAb.

Microglia Proliferation Module Antibody Sampler Kit: Image 41

Immunohistochemical analysis of paraffin-embedded mouse small intestine using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 42

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Survivin siRNA II (+), using Survivin (71G4B7E) Rabbit mAb #2808 and α-Tubulin (11H10) Rabbit mAb #2125. Survivin (71G4B7E) Rabbit mAb confirms silencing of survivin expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of survivin siRNA.

Microglia Proliferation Module Antibody Sampler Kit: Image 43

Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor (top left), 4T1 syngeneic mammary tumor (top right), Renca cell pellet (bottom left), and 4T1 cell pellet (bottom right) using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific). Both tumors show staining of infiltrating immune cells. Note the presence of staining in the Renca tumor cells and the lack of staining in the 4T1 tumor cells consistent with staining results on corresponding cell pellets.

Microglia Proliferation Module Antibody Sampler Kit: Image 44

Immunoprecipitation of ASC/TMS1 from J774A.1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is ASC (D2W8U) Rabbit mAb (Mouse Specific). Western blot analysis was performed using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific).

Microglia Proliferation Module Antibody Sampler Kit: Image 45

Western blot analysis of extracts from J774A.1 and Raw 264.7 cells using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

To Purchase # 49938T
Product # Size Price
49938T
1 Kit  (7 x 20 µl) N/A

Product Description

The Microglia Proliferation Module Antibody Sampler Kit provides an economical means of detecting proteins identified as markers of microglial proliferation by western blot and/or immunofluorescence.

Specificity / Sensitivity

Each antibody in the Microglia Proliferation Module Antibody Sampler Kit detects endogenous levels of its target protein. HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) does not recognize human HS1 protein. HS1 has a calculated size of 54 kDa, but has an apparent molecular weight of 80 kDa on SDS-PAGE gels. Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) XP® Rabbit mAb detects endogenous levels of Aurora A/B/C only when phosphorylated at either Thr288, Thr232, or Thr198 respectively. HP1α/β (C7F11) Rabbit mAb does not cross-react with HP1 γ proteins.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu310 of mouse HS1, Cys60 of human Survivin, Thr232 of human Aurora B, the carboxy terminus of human HP1α, the amino terminus of human Ki-67 and MCM2, and recombinant mouse ASC/TMSI protein.

Background

Distinct microglial activation states have been identified using RNA-seq data from a vast array of neurological disease and aging models. These activation states have been categorized into modules corresponding to proliferation, neurodegeneration, interferon-relation, LPS-relation, and many others (1). Previous work identifying markers of specific brain cell types using RNA-seq has shown HS1 and ASC/TMS1 to be useful and specific tools to study microglia (2). HS1 is a protein kinase substrate that is expressed only in tissues and cells of hematopoietic origin (3) and ASC/TMS1 has been found to be a critical component of inflammatory signaling where it associates with and activates caspase-1 in response to pro-inflammatory signals (4).

Ki-67 is a nuclear nonhistone protein (5) universally expressed among proliferating cells and absent in quiescent cells (6). Minichromosome maintenance protein 2 (MCM2) is a nuclear protein that plays a role in DNA replication and cell division (7) and is commonly used as a marker for cell proliferation, including brain tissue (8). Survivin binds and inhibits caspase-3, controlling the checkpoint in the G2/M-phase of the cell cycle by inhibiting apoptosis and promoting cell division (9). Aurora A, B, and C are a family of highly conserved serine/threonine kinases that regulate chromosomal alignment and segregation during mitosis and meiosis. Their activity requires autophosphorylation of a threonine within their kinase domain at site Thr288 of Aurora A, Thr232 of Aurora B, and Thr198 of Aurora C (10). Heterochromatin protein 1 (HP1) α and β are heterochromatic adaptor molecules involved in both gene silencing and higher order chromatin structure (11).

  1. Friedman, B.A. et al. (2018) Cell Rep 22, 832-47.
  2. Zhang, Y. et al. (2014) J Neurosci 34, 11929-47.
  3. Kitamura, D. et al. (1995) Biochem Biophys Res Commun 208, 1137-46.
  4. Srinivasula, S.M. et al. (2002) J Biol Chem 277, 21119-22.
  5. Gerdes, J. et al. (1983) Int J Cancer 31, 13-20.
  6. Weigel, M.T. and Dowsett, M. (2010) Endocr Relat Cancer 17, R245-62.
  7. Mincheva, A. et al. (1994) Cytogenet Cell Genet 65, 276-7.
  8. Doorn, K.J. et al. (2014) Neural Plast 2014, 959154.
  9. Li, F. et al. (1999) Nat Cell Biol 1, 461-6.
  10. Goldenson, B. and Crispino, J.D. (2015) Oncogene 34, 537-45.
  11. Maison, C. and Almouzni, G. (2004) Nat. Rev. Mol. Cell Biol. 5, 296-304.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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