Revision 5

#50748Store at -20C

1 Kit

(7 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
GFAP (E4L7M) XP® Rabbit mAb 80788 20 µl 50 kDa Rabbit IgG
EAAT1 (D44E2) XP® Rabbit mAb 5684 20 µl 58 kDa Rabbit IgG
Survivin (71G4B7) Rabbit mAb 2808 20 µl 16 kDa Rabbit IgG
EAAT2 (E3P5K) Rabbit mAb 20848 20 µl 65 kDa Rabbit IgG
ALDH1L1 (E7I2Q) Rabbit mAb 85828 20 µl 98 kDa Rabbit IgG
AQP4 (D1F8E) XP® Rabbit mAb 59678 20 µl 28 kDa Rabbit IgG
GAT1 (E7J1B) Rabbit mAb 37342 20 µl 65 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Astrocyte Markers Antibody Sampler Kit provides an economical means of detecting astrocyte markers by Immunofluorescence or Western Blot. The kit includes enough antibodies to perform at least two western blot or twenty IF tests with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

Astrocytes are a population of cells with distinctive morphological and functional characteristics that differ within specific areas of the brain. Postnatally, astrocyte progenitors migrate to reach their brain area and related properties. They have a regulatory role in brain functions that are implicated in neurogenesis and synaptogenesis, controlling blood-brain barrier permeability and maintaining extracellular homeostasis. Mature astrocytes also express some genes enriched in cell progenitors, suggesting they can retain proliferative potential (1). Astrocytes in the human brain are characterized by a heavy expression of the glial fibrillary acidic protein (GFAP), comprised of interlaminar that are located in layers I and II, protoplasmic in layers III and IV, varicose projections in layers V and VI, and fibrous astroglia in white matter (2,3).

Aquaporins (AQP) are integral membrane proteins that serve as channels in the transfer of water and small solutes across the membrane. AQP4 is present in the brain and it is enriched in astrocytes to regulate water homeostasis, preventing cerebral edema caused by solute imbalance (4,5). Excitatory amino acid transporters are members of sodium-dependent, high-affinity transporters that mediate the uptake of L-glutamate and D-aspartate (6). GABA transmitters are Na+/Cl--dependent transporters that regulate neurotransmitter transport, including GAT1 (SLC6), GAT2, GAT3, and BGT1 (7). GAT1 expresses in the brain, preferentially in glial cells, but is also found in neurons, regulating the uptake and release of neurotransmitters in terminal clefts (8-10). EAAT2, also known as GLT-1, is primarily expressed in astrocytes accounting for up to 90% of the total glutamate transport in the brain. EAAT1 upregulates increased concentrations of glutamate in astrocytes and has a neuroprotective potential following ischemia since reactive astrocytes and activated microglia express EAAT1 but not EAAT2 (11-13).

Aldehyde dehydrogenase 1 family member L1 (ALDH1L1) is a member of the aldehyde dehydrogenase super family (14,15). ALDH1L1 has also been shown to be a useful astrocyte marker throughout the grey and white matter of the brain, labeling both the cell body and processes of astrocytes. ALDH1L1 does not label neurons or oligodendrocytes (16). It has been shown that expression of ALDH1L1 in astrocytes is stably expressed through normal tissue and during astrogliosis (17). Survivin is a 16 kDa anti-apoptotic protein highly expressed during fetal development and cancer malignancy. It binds and inhibits caspase-3, controlling the checkpoint in the G2/M-phase of the cell cycle by inhibiting apoptosis and promoting cell division (18). Survivin expression is associated with malignant phenotypes and prognosis of glioma (19).

  1. Siracusa, R. et al. (2019) Front Pharmacol 10, 1114.
  2. Vasile, F. et al. (2017) Brain Struct Funct 222, 2017-2029.
  3. Eng, L.F. et al. (2000) Neurochem Res 25, 1439-51.
  4. Takata, K. et al. (2004) Prog Histochem Cytochem 39, 1-83.
  5. Kobayashi, H. et al. (2004) J Pharmacol Sci 96, 264-70.
  6. Amara, S.G. and Fontana, A.C. (2002) Neurochem Int 41, 313-8.
  7. Kristensen, A.S. et al. (2011) Pharmacol Rev 63, 585-640.
  8. Borden, L.A. (1996) Neurochem Int 29, 335-56.
  9. Moldavan, M. et al. (2017) J Neurophysiol 118, 3092-3106.
  10. Lorenz-Guertin, J.M. and Jacob, T.C. (2018) Dev Neurobiol 78, 238-270.
  11. Hediger, M.A. (1999) Am J Physiol 277, F487-92.
  12. Gegelashvili, G. et al. (1996) Neuroreport 8, 261-5.
  13. Beschorner, R. et al. (2007) Histopathology 50, 897-910.
  14. Krupenko, S.A. (2009) Chem Biol Interact 178, 84-93.
  15. Krupenko, S.A. and Oleinik, N.V. (2002) Cell Growth Differ 13, 227-36.
  16. Cahoy, J.D. et al. (2008) J Neurosci 28, 264-78.
  17. Michalovicz, L.T. et al. (2019) J Neurochem 150, 420-440.
  18. Reed, J.C. and Reed, S.I. (1999) Nat Cell Biol 1, E199-200.
  19. Tong, X. et al. (2019) Oncol Lett 18, 359-367.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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    Revision 5
    #50748

    Astrocyte Markers Antibody Sampler Kit

    Astrocyte Markers Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (1.0 mg/mL) from Jurkat cells using Survivin (71G4B7) Rabbit mAb #2808. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Astrocyte Markers Antibody Sampler Kit: Image 2 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal cynomolgus monkey brain using GFAP (E4L7M) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 3 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal cynomolgus monkey brain using GFAP (E4L7M) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 4 Expand Image
    Confocal immunofluorescent analysis of U-251 MG cells (left, positive) or HeLa cells (right, negative) using GFAP (E4L7M) XP® Rabbit mAb #80788 (green). Actin filaments were labeled with DyLight 650 Phalloidin #12956 (red). Blue = DAPI #4083 (fluorescent DNA dye).
    Astrocyte Markers Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from mouse brain, rat brain, and human brain using EAAT2 (E3P5K) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
    Astrocyte Markers Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of lysates from Raji (human), BaF3 (mouse) and C6 (rat) cell lines using Survivin (71G4B7E) Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various tissues using GAT1 (E7J1B) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
    Astrocyte Markers Antibody Sampler Kit: Image 8 Expand Image
    Confocal immunofluorescent analysis of rat brain using EAAT1 (D44E2) XP® Rabbit mAb (green). Red = Propidium Iodide (fluorescent DNA dye).
    Astrocyte Markers Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of mouse, rat, and human brain extracts using AQP4 (D1F8E) XP® Rabbit mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower).
    Astrocyte Markers Antibody Sampler Kit: Image 10 Expand Image
    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.
    Astrocyte Markers Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from various tissues and cell lines using GFAP (E4L7M) XP® Rabbit mAb and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Astrocyte Markers Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human brain using GFAP (E4L7M) XP® Rabbit mAb performed on the Leica BOND Rx.
    Astrocyte Markers Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from rat brain and human cortex tissue using ALDH1L1 (E7I2Q) Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of mouse retina using EAAT2 (E3P5K) Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 15 Expand Image
    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.
    Astrocyte Markers Antibody Sampler Kit: Image 16 Expand Image
    Immunoprecipitation of GAT1 protein from mouse brain lysates. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is GAT1 (E7J1B) Rabbit mAb. Western blot analysis was performed using GAT1 (E7J1B) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    Astrocyte Markers Antibody Sampler Kit: Image 17 Expand Image
    Immunoprecipitation of AQP4 from human cortex extracts. Lane 1 is human cortex lysate immunoprecipitation, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, lanes 3 and 4 represent 1:50 and 1:100 dilutions of the immunoprecipitation of human cortex lysates. Western blot analysis was performed using AQP4 (D1F8E) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded human medulloblastoma using GFAP (E4L7M) XP® Rabbit mAb performed on the Leica BOND Rx.
    Astrocyte Markers Antibody Sampler Kit: Image 19 Expand Image
    Immunoprecipitation of ALDH1L1 protein from mouse brain tissue extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is ALDH1L1 (E7I2Q) Rabbit mAb. Western blot analysis was performed using ALDH1L1 (E7I2Q) Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 20 Expand Image
    Confocal immunofluorescent analysis of mouse hindbrain using EAAT2 (E3P5K) Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded human transitional epithelial carcinoma of the bladder using Survivin (71G4B7E) Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 22 Expand Image
    Confocal immunofluorescent analysis of rat cerebellum using GAT1 (E7J1B) Rabbit mAb (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human cerebellum using AQP4 (D1F8E) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 24 Expand Image
    Confocal immunofluorescent analysis of mouse kidney (left), liver (center), and testis (right) using ALDH1L1 (E7I2Q) Rabbit mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 25 Expand Image
    Confocal immunofluorescent analysis of mouse cerebellum using EAAT2 (E3P5K) Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma showing nuclear localization using Survivin (71G4B7E) Rabbit mAb #2808.
    Astrocyte Markers Antibody Sampler Kit: Image 27 Expand Image
    Confocal immunofluorescent analysis of rat retina using GAT1 (E7J1B) Rabbit mAb (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 28 Expand Image
    Confocal immunofluorescent analysis of SW837 cells (left, positive) and A549 cells (right, negative) using EAAT1 (D44E2) XP® Rabbit mAb (green) and DAPI #4083 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using AQP4 (D1F8E) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded human glioblastoma using GFAP (E4L7M) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 31 Expand Image
    Confocal immunofluorescent analysis of mouse hippocampus using ALDH1L1 (E7I2Q) Rabbit mAb (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 647 Conjugate) #3657 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 32 Expand Image
    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).
    Astrocyte Markers Antibody Sampler Kit: Image 33 Expand Image
    Confocal immunofluorescent analysis of rat hippocampus (left) and striatum (right) using GAT1 (E7J1B) Rabbit mAb (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain near third ventricle using AQP4 (D1F8E) XP® Rabbit mAb (left) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    Astrocyte Markers Antibody Sampler Kit: Image 35 Expand Image
    Immunohistochemical analysis of paraffin-embedded human medulloblastoma using GFAP (E4L7M) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded human pituitary adenoma using Survivin (71G4B7E) Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 37 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse cerebellum using AQP4 (D1F8E) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse hippocampus using GFAP (E4L7M) XP® Rabbit mAb (left) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    Astrocyte Markers Antibody Sampler Kit: Image 39 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse kidney (left, positive) and mouse prostate (right, negative) using AQP4 (D1F8E) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 40 Expand Image
    Immunohistochemical analysis of various paraffin-embedded normal mouse tissues: brain cortex (top-left), colon (top-center), small intestine (top-right), liver (bottom-left), prostate (bottom-center) and skeletal muscle (bottom-right) using GFAP (E4L7M) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 41 Expand Image
    Immunohistochemical analysis of paraffin-embedded rat brain, cortex (left) and cerebellum (right), using GFAP (E4L7M) XP® Rabbit mAb.
    Astrocyte Markers Antibody Sampler Kit: Image 42 Expand Image
    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).
    Astrocyte Markers Antibody Sampler Kit: Image 43 Expand Image
    Confocal immunofluorescent analysis of mouse retina (left) and mouse brain (right) using AQP4 (D1F8E) XP® Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 44 Expand Image
    Confocal immunofluorescent analysis of adult mouse cerebellum (left) and hippocampus (right) using GFAP (E4L7M) XP® Rabbit mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 45 Expand Image
    Flow cytometric analysis of untreated Jurkat cells using Survivin (71G4B7E) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
    Astrocyte Markers Antibody Sampler Kit: Image 46 Expand Image
    Confocal immunofluorescent analysis of mouse kidney using AQP4 (D1F8E) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Astrocyte Markers Antibody Sampler Kit: Image 47 Expand Image
    Confocal immunofluorescent analysis of mouse colon using AQP4 (D1F8E) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).