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36833
Late-Onset Alzheimer's Disease Risk Gene Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Late-Onset Alzheimer's Disease Risk Gene Antibody Sampler Kit #36833

Citations (0)
Confocal immunofluorescent analysis of fixed frozen hippocampus from C57BL/6 (left) and APOE4 knock-in (hu/hu, model #1549-F, right) mice labeled with ApoE (pan) (D7I9N) Rabbit mAb (top, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to colabeling with GFAP (GA5) Mouse mAb (Alexa Fluor® 594 Conjugate) #8152 (bottom, red), and AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #89851 (bottom, blue pseudocolor). Mice from Taconic Biosciences, Inc.

Simple Western™ analysis of lysates (0.1 mg/mL) from 293T cells transfected with constructs expressing full-length human ApoE4 protein using ApoE4 (E5M4L) Rabbit mAb #39327. 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 2-40 kDa separation module.

Flow cytometric analysis of fixed/permeablized Jurkat cells (blue, negative) and THP-1 cells (green, positive) using TREM2 (D8I4C) Rabbit mAb (solid lines) or concentration-matached Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Western blot analysis of cell extracts from Hep G2 cells treated with Brefeldin A #9972 (10 ng/ml, 90 min) and human cerebellum using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human kidney using ApoE (pan) (D7I9N) Rabbit mAb.
Western blot analysis of extracts from indicated human cell lines and human tonsil using Clusterin (D7N2K) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of recombinant ApoE2, ApoE4, and ApoE3 proteins (5ng) using ApoE4 (E5M4L) Rabbit mAb (upper) and ApoE (pan) (D7I9N) Rabbit mAb #13366 (lower).
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using ApoE4 (E5M4L) Rabbit mAb.
Western blot analysis of extracts from various cells lines using MEF2C (D80C1) XP® Rabbit mAb.
Western blot analysis of extracts from various cells and tissue extracts using BIN1 (E4A1P) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
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).
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.
Western blot analysis of extracts from various tissues using SORL1 (D8D4G) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from LoVo, HT-29 and mIMCD-3 cell lines using EphA1 (D6V7I) Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc-tagged full-length human TREM2 protein (hTREM2-Myc; +), using TREM2 (D8I4C) Rabbit mAb.
Western blot analysis of cell extracts from 293T cells, treated with Brefeldin A #9972 (10 ng/ml, 90 min; +) and mock transfected (-) or transfected with a construct expressing full-length human ApoE2 (hApoE2; +), ApoE3 (hApoE3; +), or ApoE4 (hApoE4; +), using ApoE (pan) (D7I9N) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Immunohistochemical analysis of paraffin-embedded liver (left) or brain (right) from C57BL/6NTac (wt/wt, model #B6-F, top) and APOE4 knock-in (hu/hu, model #1549-F, bottom) mice using ApoE (pan) (D7I9N) Rabbit mAb. Mice from Taconic Biosciences, Inc.
Immunohistochemical analysis of paraffin-embedded T-47D cell pellet (left, positive) or Jurkat cell pellet (right, negative) using Clusterin (D7N2K) XP® Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing full-length human ApoE4 protein (hApoE4; +), full-length human ApoE3 protein (hApoE3; +), or full-length human ApoE2 protein (hApoE2; +), using ApoE4 (E5M4L) Rabbit mAb (upper), ApoE (pan) (D7I9N) Rabbit mAb #13366 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using ApoE4 (E5M4L) Rabbit mAb.
Confocal immunofluorescent analysis of C2C12 cells, undifferentiated (left) or differentiated for 3 days (right), using MEF2C (D80C1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Immunoprecipitation of BIN1 protein from SK-OV-3 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is BIN1 (E4A1P) Rabbit mAb. Western blot analysis was performed using BIN1 (E4A1P) Rabbit mAb.
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 THP-1, HL-60, and Jurkat cells using TREM2 (D8I4C) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation of ApoE from Hep G2 cells treated with Brefeldin A #9972 (10 ng/ml, 90 min), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or ApoE (pan) (D7I9N) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human spleen using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Clusterin (D7N2K) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded liver (left) or brain (right) from C57BL/6NTac (wt/wt, model #B6-F, top) and APOE4 knock-in (hu/hu, model #1549-F, bottom) mice using ApoE4 (E5M4L) Rabbit mAb. Mice from Taconic Biosciences, Inc.
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using MHC Class II (LGII-612.14) Mouse mAb performed on the Leica® BOND Rx.
Immunoprecipitation of TREM2 from THP-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TREM2 (D8I4C) Rabbit mAb. Western blot analysis was performed using TREM2 (D8I4C) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human small intestine using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using Clusterin (D7N2K) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human lung using ApoE4 (E5M4L) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human gastric carcinoma using MHC Class II (LGII-612.14) Mouse mAb performed on the Leica® BOND Rx.
Confocal immunofluorescent analysis of THP-1 (positive, left) and HL-60 (negative, right) cells using TREM2 (D8I4C) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human skin using ApoE (pan) (D7I9N) Rabbit mAb in the presence of control peptide (left) and antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded human tonsil using Clusterin (D7N2K) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded human mature ovarian teratoma using ApoE4 (E5M4L) 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 293T cell pellets, control (left-top) or transfected with human ApoE isoforms ApoE2 (right-top), ApoE3 (left-bottom), and ApoE4 (right-bottom) using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Clusterin (D7N2K) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using ApoE4 (E5M4L) 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 Hep G2 cell pellet (left, positive) or 293T cell pellet (right, negative) using ApoE (pan) (D7I9N) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded 293T cell pellets, control or transfected with human ApoE isoforms ApoE4, ApoE2, and ApoE3 (from left to right as indicated), using ApoE4 (E5M4L) Rabbit mAb (top panels) or ApoE (pan) (D7I9N) Rabbit mAb #13366 (bottom panels). Note the limited cross-reactivity with overexpressed ApoE2 and ApoE3. Cross-reactivity with these isoforms in endogenous models is unlikely.
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using MHC Class II (LGII-612.14) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin lymphoma using MHC Class II (LGII-612.14) Mouse mAb.
Confocal immunofluorescent analysis of Hep G2 cells (left, positive) and 293T cells (right, negative) using ApoE (pan) (D7I9N) Rabbit mAb #13366 (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
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 HepG2 cells (lane 1), 293T mock transfected (lane 2) or transiently transfected with a construct expressing ApoE4 (lane 3), whole liver extracts from wild type C57BL/6NTac model #B6-F mice (lane 4), or ApoE4 knock-in (model #1549-F) (lane 5) using ApoE4 (E5M4L) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Mice from Taconic Biosciences, Inc.
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.
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.
Western blot analysis of extracts from HepG2 cells (lane 1), 293T mock transfected (lane 2) or transiently transfected with a construct expressing ApoE4 (lane 3), whole liver extracts from wild type C57BL/6NTac model #B6-F mice (lane 4), or ApoE4 knock-in (model #1549-F) (lane 5), whole brain extracts from wild type C57BL/6NTac mice (lane 6), or ApoE4 knock-in (lane 7) using ApoE (pan) (D7I9N) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Mice from Taconic Biosciences, Inc.
To Purchase # 36833
Cat. # Size Qty. Price
36833T
1 Kit  (9 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
BIN1 (E4A1P) Rabbit mAb 51844 20 µl
  • WB
  • IP
H M 45-80 Rabbit IgG
SORL1 (D8D4G) Rabbit mAb 79322 20 µl
  • WB
H M 250 Rabbit IgG
TREM2 (D8I4C) Rabbit mAb 91068 20 µl
  • WB
  • IP
  • IF
  • F
H 28 Rabbit IgG
ApoE (pan) (D7I9N) Rabbit mAb 13366 20 µl
  • WB
  • IP
  • IHC
  • IF
H 35 Rabbit IgG
Clusterin (D7N2K) XP® Rabbit mAb 34642 20 µl
  • WB
  • IHC
H 35-42, 65, 75 Rabbit IgG
ApoE4 (E5M4L) Rabbit mAb 39327 20 µl
  • WB
  • IHC
H 35 Rabbit IgG
EphA1 (D6V7I) Rabbit mAb 90673 20 µl
  • WB
H M 130 Rabbit IgG
MEF2C (D80C1) XP® Rabbit mAb 5030 20 µl
  • WB
  • IP
  • IF
H M 50-60 Rabbit IgG
MHC Class II (LGII-612.14) Mouse mAb 68258 20 µl
  • WB
  • IP
  • IHC
  • F
H 25-35, 50-65 Mouse IgG1
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Late-Onset Alzheimer's Disease Risk Gene Antibody Sampler Kit provides an economical means of detecting proteins identified as risk factors for late-onset Alzheimer’s Disease (LOAD) by western blot. This kit includes enough antibodies to perform at least two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Late-Onset Alzheimer's Disease Risk Gene Antibody Sampler Kit detects endogenous levels of its target protein. SORL1 (D8D4G) Rabbit mAb recognizes endogenous levels of total SORL1 protein. TREM2 (D8I4C) Rabbit mAb recognizes endogenous levels of total TREM2 protein. Clusterin (D7N2K) XP® Rabbit mAb recognizes endogenous levels of total Clusterin protein. EphA1 (D6V7I) Rabbit mAb recognizes endogenous levels of total EphA1 protein. MEF2C (D80C1) XP® Rabbit mAb detects endogenous levels of total MEF2C protein. BIN1 (E4A1P) Rabbit mAb recognizes endogenous levels of total BIN1 protein. The antibody recognizes multiple BIN1 isoforms. ApoE (pan) (D7I9N) Rabbit mAb recognizes endogenous levels of total ApoE protein. This antibody also recognizes overexpressed ApoE2, ApoE3, and ApoE4 proteins. ApoE4 (E5M4L) Rabbit mAb recognizes endogenous levels of total ApoE4 protein. This antibody does not cross-react with ApoE2 or ApoE3 by western blot and is not expected to cross-react with endogenous ApoE2 or ApoE3 by immunohistochemistry. Non-specific staining was observed in kidney by immunohistochemistry. 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, or 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 a synthetic peptide corresponding to residues surrounding Val266 of human BIN1 protein, Glu267 of human SORL1 protein, Leu221 of human TREM2 protein, Pro285 of human ApoE protein, Ser396 of human Clusterin protein, Arg130 of human ApoE4 protein, Met182 of human MEF2 protein, and a recombinant protein fragment specific to the extracellular domain of human EphA1 protein. MHC Class II (LGII-612.14) Mouse mAb is produced by immunizing animals with cultured human B lymphoid cells treated with IFN-gamma.

Background

Alzheimer's Disease (AD) is the leading cause of dementia worldwide. Clinically, it is characterized by the presence of extracellular amyloid plaques and intracellular neurofibrillary tangles, which result in neuronal dysfunction and cell death (1). Genome-wide association studies (GWAS) have identified a cohort of risk genes associated with late-onset AD (LOAD), including, but not limited to, APOE, BIN1, SORL1, TREM2, EphA1, MEF2C, CLU, and HLA-DRB1 (2,3).
 
APOE has three allele variants: ApoE2, ApoE3, and ApoE4. ApoE4 is associated with an increased risk of AD. Evidence suggests that this risk occurs through promotion of amyloid-beta plaque aggregation (1). ApoE4 is also associated with impaired microglial response, lipid transport, synaptic integrity and plasticity, glucose metabolism, and cerebrovascular integrity (4). Mutations in BIN1, primarily involved in endocytosis and maintaining cytoskeletal integrity in the brain, are suggested to play a role in the aggravation of tau pathology (5,6). Increased levels of BIN1 have been seen in AD postmortem brain tissue (6). SORL1 expression is decreased in the brain of AD patients (7). Studies have demonstrated a role for SORL1 as a neuronal sorting receptor that binds amyloid precursor protein (APP) and regulates its trafficking and proteolytic processing, thus regulating β-amyloid (Aβ) peptide production (8). The triggering receptor expressed on myeloid cells 2 (TREM2) is an innate immune receptor that is expressed on the cell surface of microglia, macrophages, osteoclasts, and immature dendritic cells (9). Research studies using AD mouse models indicate that deficiency and haploinsufficiency of TREM2 can lead to increased Aβ accumulation due to dysfunctional microglia response (10). EphA1 is a member of the ephrin family of receptor tyrosine kinases responsible for regulating cell morphology and motility (11). In the central nervous system (CNS), EphA1 plays a role in synaptic plasticity and axon guidance (12). EphA1 is involved in inflammatory signaling pathways (13), which may mean it plays a role in regulation of neuroinflammatory processes in AD (14). MEF2C is a member of the myocyte enhancer factor 2 (MEF2) family of transcription factors shown to play a role in learning and memory formation through regulation of synaptic plasticity (15). Studies have shown that MEF2C may play a role in age-related microglial activation through IFN-I associated MEF2C deregulation (16,17). MEF2C may also act as a modulator for APP proteolytic processing of Aβ (18,19). Clusterin (CLU) is a multifunctional glycoprotein shown to play a protective role in AD by sequestering Aβ40 peptides to form long-lived, stable complexes, which prevent amyloid fibril formation (20-22). Major histocompatibility complex class II (MHC class II) molecules are transmembrane glycoproteins expressed on the surface of antigen-presenting cells that bind exogenous peptide antigens derived from endocytosed extracellular proteins digested in the lysosome (23,24). Increases in MHC class II-expressing microglia have been shown in AD brain (25).

  1. Selkoe, D.J. (2001) Physiol Rev 81, 741-66.
  2. Jansen, I.E. et al. (2019) Nat Genet 51, 404-413.
  3. Zhang, Q. et al. (2020) Nat Commun 11, 4799.
  4. Yamazaki, Y. et al. (2019) Nat Rev Neurol 15, 501-518.
  5. Franzmeier, N. et al. (2019) Nat Commun 10, 1766.
  6. Chapuis, J. et al. (2013) Mol Psychiatry 18, 1225-34.
  7. Scherzer, C.R. et al. (2004) Arch Neurol 61, 1200-5.
  8. Andersen, O.M. et al. (2005) Proc Natl Acad Sci U S A 102, 13461-6.
  9. Colonna, M. (2003) Nat Rev Immunol 3, 445-53.
  10. Wang, Y. et al. (2015) Cell 160, 1061-71.
  11. Yamazaki, T. et al. (2009) J Cell Sci 122, 243-55.
  12. Lai, K.O. and Ip, N.Y. (2009) Curr Opin Neurobiol 19, 275-83.
  13. Ivanov, A.I. and Romanovsky, A.A. (2006) IUBMB Life 58, 389-94.
  14. Villegas-Llerena, C. et al. (2016) Curr Opin Neurobiol 36, 74-81.
  15. Rashid, A.J. et al. (2014) Genes Brain Behav 13, 118-25.
  16. Xue, F. et al. (2021) Neurobiol Dis 152, 105272.
  17. Deczkowska, A. et al. (2017) Nat Commun 8, 717.
  18. Tang, S.S. et al. (2016) Oncotarget 7, 39136-39142.
  19. Camargo, L.M. et al. (2015) PLoS One 10, e0115369.
  20. Yerbury, J.J. et al. (2007) FASEB J 21, 2312-22.
  21. Narayan, P. et al. (2011) Nat Struct Mol Biol 19, 79-83.
  22. Desikan, R.S. et al. (2014) JAMA Neurol 71, 180-7.
  23. Ting, J.P. and Trowsdale, J. (2002) Cell 109 Suppl, S21-33.
  24. Cresswell, P. (1994) Annu Rev Immunol 12, 259-93.
  25. Perlmutter, L.S. et al. (1992) J Neurosci Res 33, 549-58.

Pathways

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