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78646
Genetics of Parkinson's Disease: Lysosomal Dysfunction Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Genetics of Parkinson's Disease: Lysosomal Dysfunction Antibody Sampler Kit #78646

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Simple Western™ analysis of extracts (1 mg/mL) from Mouse brain tissue using α-Synuclein (E4U2F) XP® Rabbit mAb #51510. The virtual lane view (left) shows a single 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 12-230 kDa separation module.
Simple Western™ analysis of lysates (1 mg/mL) from PC-12 cells using GCase/GBA (E2R1L) Rabbit mAb #88162. 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 12-230 kDa separation module.
Western blot analysis of extracts from U-87 MG and A172 cells, and mouse brain using LRRK2 (D18E12) Rabbit mAb. 
Western blot analysis of extracts from mouse and rat brain using α-Synuclein (E4U2F) XP® Rabbit mAb.
Immunoprecipitation of α-Synuclein 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 α-Synuclein (E4U2F) XP® Rabbit mAb. Western blot analysis was performed using α-Synuclein (D37A6) Rabbit mAb #4179. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Confocal immunofluorescent analysis of fixed frozen mouse cerebellum labeled with α-Synuclein (E4U2F) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb #71299 (right, red) and DAPI #4083 (right, blue).
Confocal immunofluorescent analysis of fixed frozen mouse striatum labeled with α-Synuclein (E4U2F) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb #71299 (right, red), and DAPI #4083 (right, blue).
Confocal immunofluorescent analysis of SK-MEL-28 cells (left, positive) and HuH-6 cells (right, negative) using α-Synuclein (E4U2F) XP® Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Immunohistochemical analysis of paraffin-embedded normal human brain using α-Synuclein (E4U2F) XP® Rabbit mAb.
Western blot analysis of extracts from HeLa and COS-7 cells using PARK9 Antibody.
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 brain tissues and HCC1419 cells using Cathepsin D (E5V4H) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Confocal immunofluorescent analysis of fixed frozen brain from an amyloid mouse model of Alzheimer's disease using Cathepsin D (E5V4H) Rabbit mAb (left, green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #36618 (right, red) and β-Amyloid (D3D2N) Mouse mAb #15126 (right, blue).
Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the cervix using Cathepsin D (E5V4H) Rabbit mAb.
Western blot analysis of extracts from various cell lines using VPS35 (E6S4I) Rabbit mAb.
Western blot analysis of extracts from immortalized gba+/+ and gba-/- mouse neurons using GCase/GBA (E2R1L) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Immortalized gba+/+ and gba-/- mouse neurons kindly provided by Dr. Ellen Sidransky, MD at the National Human Genome Research Institute of the National Institutes of Health (8).
Western blot analysis of A549 extracts from WT (left) or LRRK2 KO (right) using LRRK2 (D18E12) Rabbit mAb. Membranes stained with Ponceau S for total protein normalization (lower). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies, as a companion to validation data generated by CST scientists.
Immunohistochemical analysis of paraffin-embedded mouse cerebellum using α-Synuclein (E4U2F) XP® Rabbit mAb.
Confocal immunofluorescent analysis of fixed frozen mouse brain using Cathepsin D (E5V4H) Rabbit mAb (left, green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #36618 (right, red) and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunohistochemical analysis of paraffin-embedded human papillary thyroid carcinoma using Cathepsin D (E5V4H) Rabbit mAb.
Immunoprecipitation of VPS35 protein from U-251 MG cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is VPS35 (E6S4I) Rabbit mAb. Western blot analysis was performed using VPS35 (E6S4I) Rabbit mAb.
Western blot analysis of extracts from various cell lines using GCase/GBA (E2R1L) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Simple WesternTM analysis of lysates (1.0 mg/mL) from Mouse Brain cells using LRRK2 (D18E12) Rabbit mAb #13046. 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 JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
Immunohistochemical analysis of paraffin-embedded normal human spleen using α-Synuclein (E4U2F) XP® Rabbit mAb.
Confocal immunofluorescent analysis of fixed frozen mouse colon at high magnification using Cathepsin D (E5V4H) Rabbit mAb (left, green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #36618 (right, red) and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using Cathepsin D (E5V4H) Rabbit mAb.
Immunoprecipitation of LRRK2 from mouse brain extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is LRRK2 (D18E12) Rabbit mAb. Western blot analysis was performed using LRRK2 (D18E12) Rabbit mAb. Anti-rabbit, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded normal human kidney using α-Synuclein (E4U2F) XP® Rabbit mAb.
Confocal immunofluorescent analysis of fixed frozen mouse colon at low magnification using Cathepsin D (E5V4H) Rabbit mAb (left, green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #36618 (right, red) and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunohistochemical analysis of paraffin-embedded normal human adrenal gland using Cathepsin D (E5V4H) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using α-Synuclein (E4U2F) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HCC1419 cells (left) and U-118 MG cells (right) using Cathepsin D (E5V4H) Rabbit mAb (green), β-Actin (8H10D10) Mouse mAb #3700 (red), and DAPI #4083 (blue).
Immunohistochemical analysis of paraffin-embedded normal human pancreas using Cathepsin D (E5V4H) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using α-Synuclein (E4U2F) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human lymph node using Cathepsin D (E5V4H) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse spleen using α-Synuclein (E4U2F) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human brain using Cathepsin D (E5V4H) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse liver using α-Synuclein (E4U2F) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse pancreas using Cathepsin D (E5V4H) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse kidney using α-Synuclein (E4U2F) XP® Rabbit mAb.


Immunohistochemical analysis of paraffin-embedded mouse thymus using Cathepsin D (E5V4H) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using α-Synuclein (E4U2F) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).

Immunohistochemical analysis of paraffin-embedded mouse testes using Cathepsin D (E5V4H) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human normal brain (left), urothelial carcinoma (middle) or lung adenocarcinoma (right) using α-Synuclein (E4U2F) XP® Rabbit mAb (top) or α-Synuclein Mouse mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on human α-Synuclein. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining. 
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma (left) and tonsil (right) using Cathepsin D (E5V4H) Rabbit mAb (top) or a Cathepsin D Rabbit pAb (bottom). These two antibodies detect unique, non-overlapping epitopes on human cathepsin D. The similar patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemical analysis of paraffin-embedded SK-MEL-28 cell pellet (left, positive) or HuH-6 cell pellet (right, negative) using α-Synuclein (E4U2F) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse cerebellum (left) and A20 syngeneic tumor (right) using Cathepsin D (E5V4H) Rabbit mAb (top) or a Cathepsin D Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on mouse cathepsin D. The similar patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemical analysis of paraffin-embedded normal human liver using Cathepsin D (E5V4H) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
Immunohistochemical analysis of paraffin-embedded U-118 MG cell pellet (left, positive) or Ramos cell pellet (right, negative) using Cathepsin D (E5V4H) Rabbit mAb.
To Purchase # 78646
Cat. # Size Qty. Price
78646T
1 Kit  (6 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
GCase/GBA (E2R1L) Rabbit mAb 88162 20 µl
  • WB
H M R 65 Rabbit IgG
Cathepsin D (E5V4H) Rabbit mAb 74089 20 µl
  • WB
  • IHC
  • IF
H M R 46, 43, 28 Rabbit IgG
PARK9 Antibody 5879 20 µl
  • WB
  • IP
H M R Mk 150 Rabbit 
VPS35 (E6S4I) Rabbit mAb 81453 20 µl
  • WB
  • IP
H M R Mk 81 Rabbit IgG
LRRK2 (D18E12) Rabbit mAb 13046 20 µl
  • WB
  • IP
H M R 290 Rabbit IgG
α-Synuclein (E4U2F) XP® Rabbit mAb 51510 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R 18 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Genetics of Parkinson's Disease: Lysosomal Dysfunction Antibody Sampler Kit provides an economical means of investigating lysosomal proteins that are commonly mutated in Parkinson’s disease (PD) by western blot. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Background

The antibodies in this kit serve to characterize genes implicated in Parkinson’s disease (PD) that lead to lysosomal dysfunction. The cellular function of lysosomes is to degrade and recycle cellular waste, maintaining proper cellular energy metabolism. GBA mutations are the most common genetic risk factor for PD, a neurodegenerative disease characterized by the loss of dopaminergic neurons in the substantia nigra with formation of Lewy bodies in surviving neurons (1,2). In the absence of GBA, which encodes β-glucocerebrosidase (GCase), autophagic lysosome reformation is altered, suggesting that GCase activity is critical to maintaining functional lysosomes. GCase catalyzes the hydrolysis of glucocerebroside into free ceramide and glucose (3). Lysosomal breakdown of glucocerebroside is required for complex lipid cellular metabolism and proper cellular membrane turnover (4).

α-Synuclein (α-Syn) is the main component of pathogenic Lewy bodies and neurites. Research studies have shown that mutations in the α-Synuclein gene are linked to PD (5). Neuronally generated pathogenic species of α-Syn accumulate within neuronal lysosomes and are released via SNARE-dependent lysosomal exocytosis. The released aggregates are non-membrane enveloped and seeding-competent, further contributing to PD pathology (6).

PARK9, also known as ATP13A2, is a member of the P-type ATPase superfamily. It localizes to the lysosomal membrane and is involved in the lysosomal degradation pathway, clearing α-Syn aggregates (7,8).

Cathepsin D (CTSD) is a ubiquitously expressed lysosomal aspartyl protease involved in normal protein degradation. Loss of CTSD leads to lysosomal dysfunction and accumulation of different cellular proteins implicated in neurodegenerative diseases, such as α-Syn (9).

Another gene genetically linked to PD is leucine-rich repeat kinase 2 (LRRK2), also known as PARK8 (10). Research studies have linked at least 20 mutations in LRRK2 to PD, with the G2019S mutation being the most prevalent (11). LRRK2 mutations alter lysosome morphology, distribution, pH, and degradative capacity in many distinct cell types, including fibroblasts, striatal neurons, primary neurons, and primary astrocytes (12).

Mutations in the vacuolar protein sorting-associated protein 35 (VPS35) gene, encoding a core component of the retromer complex, have recently emerged as a new cause of late-onset, autosomal dominant familial PD. A single missense mutation, Asp620Asn (D620N), has so far been unambiguously identified to cause PD (13). The VPS35 D620N mutation alters the expression of ~220 lysosomal proteins and stimulates recruitment of LRRK2 and phosphorylation of Rab proteins at the lysosome (14).

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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