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41608
β-Amyloid Mouse Model Neuronal Viability IF Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

β-Amyloid Mouse Model Neuronal Viability IF Antibody Sampler Kit #41608

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Simple Western™ analysis of lysates (0.1 mg/mL) from mouse brain using PSD95 (D27E11) XP® Rabbit mAb #3450. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (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 66-440 kDa separation module.
Immunoprecipitation of cleaved caspase-3 from Jurkat extracts treated with Etoposide #2200 (25 mM; 5 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cleaved Caspase-3 (Asp175) Antibody. Western blot analysis was performed using Cleaved Caspase-3 (Asp175) Antibody. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Simple Western™ analysis of lysates (0.1 mg/mL) from Jurkat cells treated with Cytochrome C using Cleaved Caspase-3 (Asp175) Antibody #9661. The virtual lane view (left) shows two target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (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.
Western blot analysis of human Aβ-37, Aβ-40, and Aβ-42 peptides using β-Amyloid (D3D2N) Mouse mAb.
Western blot analysis of extracts from mouse brain, C2C12 cells, and rat brain using NeuN (D4G4O) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various tissues and cell lines using GFAP (E6N9L) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from human cerebellum and rat brain using PSD95 (D27E11) XP® Rabbit mAb.
Western blot analysis of cell extracts from Baf3, 32D, and mouse spleen using HS1 (D5A9) XP® Rabbit mAb.
Western blot analysis of human Aβ-42, Aβ-40, Aβ-39, Aβ-38, and Aβ-37 peptides (5 ng) using β-Amyloid (D54D2) XP® Rabbit mAb.
Confocal immunofluorescent analysis of mouse brain (left) and mouse retina (right) using Synaptophysin (7H12) Mouse mAb (IF Formulated) (green) and CNPase (D83E10) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #5715 (red, left). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye).
Western blot analysis of extracts from serum-starved Neuro-2a cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from HeLa, NIH/3T3 and C6 cells untreated, staurosporine-treated (3hrs, 1 µM in vivo) or cytochrome c-treated (1hr, 0.25 mg/ml in vitro), using Caspase-3 Antibody #9662 (upper) or Cleaved Caspase-3 (Asp175) Antibody (lower).
Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain using β-Amyloid (D3D2N) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded mouse cerebellum using NeuN (D4G4O) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded cynomolgus monkey brain using NeuN (D4G4O) XP® Rabbit mAb.
Western blot analysis of extracts from mouse brain using GFAP (E6N9L) Mouse mAb as the primary antibody. Various anti-mouse isotype specific antibodies (upper) and Anti-mouse IgG, HRP-linked Antibody #7076 (lower) were used as secondary antibodies.
Confocal immunofluorescent analysis of rat cerebellum and retina using PSD95 (D27E11) XP® Rabbit mAb (red) and Neurofilament-L (DA2) Mouse mAb #2835 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded mouse spleen using HS1 (D5A9) XP® Rabbit mAb.
Western blot analysis of the indicated amounts of human Aβ-42 (left) and Aβ-40 (right) peptides using β-Amyloid (D54D2) XP® Rabbit mAb.
Western blot analysis of extracts from serum-starved H-4-II-E cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 6 hr; +), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemical analysis of paraffin-embedded human tonsil, showing cytoplasmic and perinuclear localization in apoptotic cells (low and high magnification), using Cleaved Caspase-3 (Asp175) Antibody.
Confocal immunofluorescent analysis of brain sections from 13-month old wild-type (left) and Tg2576 (center) mice using β-Amyloid (D3D2N) Mouse mAb (green) and β3-Tubulin (D71G9) XP® Rabbit mAb #5568 (red). A detailed view of a cortical β-amyloid plaque is shown on the right. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded mouse colon (myenteric plexus) using NeuN (D4G4O) XP® Rabbit mAb.
Immunoprecipitation of GFAP protein from mouse brain tissue extracts. Lane 1 is 10% input, lane 2 is Mouse (E5Y6Q) mAb IgG2a Isotype Control #61656, and lane 3 is GFAP (E6N9L) Mouse mAb. Western blot analysis was performed GFAP (E6N9L) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded LL2 syngeneic tumor using HS1 (D5A9) XP® Rabbit mAb.
Western blot analysis of human Aβ-42 peptide (1 ng) and human cerebrospinal fluid (CSF) of an AD patient using β-Amyloid (D54D2) XP® Rabbit mAb.
Immunoprecipitation of Cleaved PARP (Asp214) from Neuro-2a cell extracts treated with Staurosporine #9953 (1 μM, 3 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cleaved PARP (Asp214) (D6X6X) Rabbit mAb. Western blot was perform using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunohistochemical analysis using Cleaved caspase-3 (Asp175) antibody on SignalSlide Cleaved Caspase-3 IHC controls #8104 (paraffin-embedded Jurkat cells, untreated (left) or etoposide-treated (right)).
Immunohistochemical analysis of paraffin-embedded human glioblastoma multiform using NeuN (D4G4O) XP® Rabbit mAb.
Confocal immunofluorescent analysis of mouse Tg2576 brain, which overexpresses mutant human APP695. Sections were labeled with GFAP (E6N9L) Mouse mAb (green, mouse IgG2a), β-Amyloid (D3D2N) Mouse mAb #15126 (red, mouse IgG1), and HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) #3892 (magenta, rabbit IgG). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Confocal immunofluorescent analysis of fixed frozen mouse cortex from wild-type (left) or an amyloid mouse model of Alzheimer's disease (right) using HS1 (D5A9) XP® Rabbit mAb (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using β-Amyloid (D54D2) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) #35363 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunohistochemical analysis of paraffin-embedded 3T3 cell pellet, untreated (left, negative) or treated with Staurosporine #9953 (right, positive), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse embryo, using Cleaved Caspase-3 (Asp175) Antibody preincubated with control peptide (left) or Cleaved Caspase-3 (Asp175) Blocking Peptide #1050 (right).
Confocal immunofluorescent analysis of mouse hippocampus (left), cortex (middle), and cerebellum (right) using NeuN (D4G4O) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of mouse Tg2576 brain which overexpresses mutant human APP695. Sections were first labeled with HS1 (D5A9) XP® Rabbit mAb (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).
Confocal immunofluorescent analysis of mouse subicular cortex from an amyloid mouse model of Alzheimer's Disease using β-Amyloid (D54D2) XP® Rabbit mAb #8243 (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunohistochemical analysis of paraffin-embedded E14 rat embryo using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb.
Confocal immunofluorescent analysis of U-251 MG cells (left, positive) or HeLa cells (right, negative) using GFAP (E6N9L) Mouse mAb #34001 (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Confocal immunofluorescent analysis of 32D cells (left) and C2C12 cells (right), using HS1 (D5A9) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded H-4-II-E cell pellet, untreated (left, negative) or treated with Staurosporine #9953 (right, positive), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb.
Confocal immunofluorescent images of HT-29 cells, untreated (left) or Staurosporine #9953 treated (right), labeled with Cleaved Caspase-3 (Asp175) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
Flow cytometric analysis of NIH/3T3 cells (blue, negative) and 32D clone 3 cells (green, positive) using HS1 (D5A9) XP® Rabbit mAb (solid lines) or 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.
Immunohistochemical analysis of paraffin-embedded mouse ovary using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb.
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with etoposide #2200 (green), using Cleaved Caspase-3 (Asp175) Antibody compared to a nonspecific negative control antibody (red).
Immunohistochemical analysis of paraffin-embedded mouse spleen using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb.
Confocal immunofluorescent analysis of Neuro-2a cells, untreated (left, negative) or treated with Staurosporine #9953 (1 μM, 3 hr; right, positive), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of serum-starved Neuro-2a cells, untreated (blue) or treated with Staurosporine #9953 (1 μM, 3 hr; green), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (solid lines) or 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.
Flow cytometric analysis of serum-starved H-4-II-E cells, untreated (blue) or treated with Staurosporine #9953 (1 μM, 3 hr; green), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (solid lines) or 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.
To Purchase # 41608
Cat. # Size Qty. Price Inventory
41608T
1 Kit  (9 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
β-Amyloid (D54D2) XP® Rabbit mAb 8243 20 µl
  • WB
  • IP
  • IF
H 5 Rabbit IgG
β-Amyloid (D3D2N) Mouse mAb 15126 20 µl
  • WB
  • IHC
  • IF
H 5 Mouse IgG1
NeuN (D4G4O) XP® Rabbit mAb 24307 20 µl
  • WB
  • IHC
  • IF
H M R Mk 46-55 Rabbit IgG
Synaptophysin (7H12) Mouse mAb (IF Formulated) 9020 20 µl
  • IF
H M R Mouse IgG1
PSD95 (D27E11) XP® Rabbit mAb 3450 20 µl
  • WB
  • IF
H M R 95 Rabbit IgG
Cleaved Caspase-3 (Asp175) Antibody 9661 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Mk 17, 19 Rabbit 
Cleaved PARP (Asp214) (D6X6X) Rabbit mAb 94885 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
M R 89 Rabbit IgG
GFAP (E6N9L) Mouse mAb 34001 20 µl
  • WB
  • IP
  • IF
H M R 50 Mouse IgG2a
HS1 (D5A9) XP® Rabbit mAb 3892 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
M 80 Rabbit IgG

Product Description

The β-Amyloid Mouse Model Neuronal Viability IF Antibody Sampler Kit provides an economical means of detecting proteins to confirm neuronal viability and surrounding astrocytes and microglia in mouse models by immunofluorescence.

Specificity / Sensitivity

Each antibody in the β-Amyloid Mouse Model Neuronal Viability IF Antibody Sampler Kit detects endogenous levels of its target protein. β-Amyloid (D54D2) XP® Rabbit mAb and β-Amyloid (D3D2N) Mouse mAb detect several isoforms of Aβ, such as Aβ-37, Aβ-38, Aβ-39, Aβ-40, and Aβ-42, and they also detect transgenically expressed human APP in mouse models. Cleaved PARP (Asp214) (D6X6X) Rabbit mAb recognizes endogenous levels of the large fragment (89 kDa) of PARP protein only when cleaved at Asp214. 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. Cleaved Caspase-3 (Asp175) Antibody detects endogenous levels of the large fragment (17/19 kDa) of activated caspase-3 resulting from cleavage adjacent to Asp175. This antibody does not recognize full length caspase-3 or other cleaved caspases. This antibody detects non-specific caspase substrates by western blot. Non-specific labeling may be observed by immunofluorescence in specific sub-types of healthy cells in fixed-frozen tissues (e.g. pancreatic alpha-cells). Nuclear background may be observed in rat and monkey samples.

Source / Purification

Monoclonal and polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of human β-amyloid peptide (Aβ), residues surrounding Gln53 of human PSD95, Asp214 of rodent PARP1, Leu310 of mouse HS1, amino-terminal residues adjacent to Asp175 of human caspase-3, recombinant protein specific to the carboxy terminus of human SYP protein and the amino terminus of human NeuN, and native GFAP purified from pig spinal cord.

Background

Amyloid β (Aβ) precursor protein (APP) is a 100-140 kDa transmembrane glycoprotein that exists as several isoforms. The amino acid sequence of APP contains the amyloid domain, which can be released by a two-step proteolytic cleavage. The extracellular deposition and accumulation of the released Aβ fragments form the main components of amyloid plaques, a major pathological hallmark of Alzheimer’s disease (1). Neuronal nuclei (NeuN, Fox-3, RBFOX3) is a nuclear protein expressed in most post-mitotic neurons of the central and peripheral nervous systems. NeuN is not detected in Purkinje cells, sympathetic ganglion cells, Cajal-Retzius cells, INL retinal cells, inferior olivary, or dentate nucleus neurons (2). Glial fibrillary acidic protein (GFAP) is the main intermediate filament in mature brain astroglial and radial glial cells and GFAP also plays an important role in modulating astroglial motility and shape (3). HS1 is a protein kinase substrate that is expressed only in tissues and cells of hematopoietic origin (4). Previous work identifying markers of specific brain cell types using RNA-seq has shown HS1 to be a useful and specific tool to study microglia (5). Synaptophysin (SYP) is a neuronal synaptic vesicle glycoprotein that occurs in presynaptic vesicles of neurons (6). Postsynaptic Density protein 95 (PSD95) is a member of the membrane-associated guanylate kinase (MAGUK) family of proteins. PSD95 is a scaffolding protein involved in the assembly and function of the postsynaptic density complex (7,8). Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins, including nuclear enzyme poly (ADP-ribose) polymerase (PARP) (9). PARP, a 116 kDa nuclear poly (ADP-ribose) polymerase, appears to be involved in DNA repair in response to environmental stress (10). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (11).

Pathways

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