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87118
Lysosomal Marker Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Lysosomal Marker Antibody Sampler Kit #87118

Citations (0)
Immunoprecipitation of mucolipin-1 protein from TT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Mucolipin-1 (F8F9Q) Rabbit mAb. Western blot analysis was performed using Mucolipin-1 (F8F9Q) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Confocal immunofluorescent analysis of TT cells (left, high-expressing) and MCF7 cells (right, low-expressing) cells using Mucolipin-1 (F8F9Q) Rabbit mAb (green), β-Actin (8H10D10) Mouse mAb #3700 (red), and DAPI #4083 (blue).
Western blot analysis of extracts from CHO-K1 cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human CD63 protein (hCD63-Myc/DDK; +), using CD63 (E1W3T) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or β-Actin Antibody #4967 (lower).
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.
Immunoprecipitation of LAMP1 from SNB-19 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is LAMP1 (D2D11) XP® Rabbit mAb. Western blot was performed using LAMP1 (D2D11) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Western blot analysis of extracts from various cell lines using Mucolipin-1 (F8F9Q) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of mucolipin-1 protein in MCF7 and Ramos cells is consistent with the predicted expression pattern.
Western blot analysis of extracts from Neuro-2a and mIMCD-3 cells, untreated (-) or treated with peptide-N-glycosidase F (PNGase F; +), using LIMP-2/SCARB2 (E2Z5F) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines using Cathepsin B (D1C7Y) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from 3T3-L1 and mIMCD-3 cells using LAMP2 (E6A6S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various human cell lines using ATP6V1A (E5N9E) Rabbit mAb.
Western blot analysis of extracts from various cell lines using CD63 (E1W3T) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of CD63 protein in Jurkat and MOLT-4 cells is consistent with the predicted expression pattern.
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 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 extracts from various cell lines using LAMP1 (D2D11) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using LIMP-2/SCARB2 (E2Z5F) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing full-length human Cathepsin B (hCTSB; +) or mouse Cathespin B (mCTSB; +) using Cathepsin B (D1C7Y) XP® Rabbit mAb.
Western blot analysis of extracts from mouse kidney, liver, and lung tissue using LAMP2 (E6A6S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from HeLa cells, transfected with control siRNA (-) or ATP6V1A siRNA (+), using ATP6V1A (E5N9E) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
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.
Western blot analysis of extracts from various cell lines using GCase/GBA (E2R1L) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemical analysis of normal human kidney using LAMP1 (D2D11) XP® Rabbit mAb.
Immunoprecipitation of LIMP-2/SCARB2 protein from Neuro-2a cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is LIMP-2/SCARB2 (E2Z5F) Rabbit mAb. Western blot analysis was performed using LIMP-2/SCARB2 (E2Z5F) Rabbit mAb. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Cathepsin B (D1C7Y) XP(R) Rabbit mAb.
Confocal immunofluorescent analysis of fixed frozen mouse intestine using LAMP2 (E6A6S) Rabbit mAb (left, green), DyLight 554 Phalloidin #13054 (right, red), and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human ATP6V1A (hATP6V1A-Myc/DDK; +), using ATP6V1A (E5N9E) Rabbit mAb (upper), DYKDDDDK Tag (D6W5B) Rabbit mAb #14793 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
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.
Immunohistochemical analysis of human lung carcinoma using LAMP1 (D2D11) XP® Rabbit mAb.
Confocal immunofluorescent analysis of fixed frozen mouse cortex at low magnification (left) and high magnification (right) using LIMP-2/SCARB2 (E2Z5F) Rabbit mAb (green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to labeling with Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #36618 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunohistochemical analysis of paraffin-embedded normal human kidney using Cathepsin B (D1C7Y) XP(R) Rabbit mAb.
Confocal immunofluorescent analysis of fixed frozen mouse kidney using LAMP2 (E6A6S) Rabbit mAb (left, green), DyLight 554 Phalloidin #13054 (right, red), and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
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 human ovarian serous cystadenoma using LAMP1 (D2D11) XP® Rabbit mAb.
Confocal immunofluorescent analysis of fixed frozen mouse small intestine (left) and kidney (right) using LIMP-2/SCARB2 (E2Z5F) Rabbit mAb (green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to labeling with Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #36618 (red), Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182 (magenta), and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Cathepsin B (D1C7Y) XP(R) Rabbit mAb.
Confocal immunofluorescent analysis of RAW 264.7 cells (left, high-expressing) or Neuro-2a cells (right, low-expressing) using LAMP2 (E6A6S) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and DAPI #4083 (blue).
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.
Confocal immunofluorescent analysis of HeLa cells using LAMP1 (D2D11) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of Neuro-2a cells (left, high-expressing) and mIMCD-3 cells (right, low-expressing) using LIMP-2/SCARB2 (E2Z5F) Rabbit mAb (green), β-Actin (8H10D10) Mouse mAb #3700 (red), and DAPI #4083 (blue).
Immunohistochemical analysis of paraffin-embedded normal human lymph node using Cathepsin D (E5V4H) Rabbit mAb.
Flow cytometric analysis of Jurkat cells using LAMP1 (D2D11) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Confocal immunofluorescent analysis of Malme-3M (left) and MCF7 (right) cells using Cathepsin (D1C7Y) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded normal human brain using Cathepsin D (E5V4H) Rabbit mAb.
Flow cytometric analysis of fixed/permeabilized Daudi cells (blue, negative) and SK-MEL-28 cells (green, positive) using Cathepsin B (D1C7Y) XP® Rabbit mAb (solid lines) or concentration-matched 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.
Immunohistochemical analysis of paraffin-embedded mouse pancreas using Cathepsin D (E5V4H) Rabbit mAb.


Immunohistochemical analysis of paraffin-embedded mouse thymus using Cathepsin D (E5V4H) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded mouse testes using Cathepsin D (E5V4H) Rabbit mAb.
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 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 # 87118
Cat. # Size Qty. Price
87118T
1 Kit  (9 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
LAMP1 (D2D11) XP® Rabbit mAb 9091 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H Mk 42 (non-glycosylated), 90-120 (glycosylated) Rabbit IgG
LAMP2 (E6A6S) Rabbit mAb 34141 20 µl
  • WB
  • IF
M 100-130 Rabbit IgG
Cathepsin B (D1C7Y) XP® Rabbit mAb 31718 20 µl
  • WB
  • IHC
  • IF
  • F
H M R 44, 27, 24 Rabbit IgG
Cathepsin D (E5V4H) Rabbit mAb 74089 20 µl
  • WB
  • IHC
  • IF
H M R 46, 43, 28 Rabbit IgG
Mucolipin-1 (F8F9Q) Rabbit mAb 27748 20 µl
  • WB
  • IP
  • IF
H 35-40, 65 Rabbit IgG
CD63 (E1W3T) Rabbit mAb 52090 20 µl
  • WB
H Mk 25-60 Rabbit IgG
GCase/GBA (E2R1L) Rabbit mAb 88162 20 µl
  • WB
H M R 65 Rabbit IgG
LIMP-2/SCARB2 (E2Z5F) Rabbit mAb 27960 20 µl
  • WB
  • IP
  • IF
H M R 54, 80 Rabbit IgG
ATP6V1A (E5N9E) Rabbit mAb 39517 20 µl
  • WB
H M R 75 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Product Description

The Lysosomal Marker Antibody Sampler Kit provides an economical means of detecting proteins localized to lysosomes. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Background

Lysosomes are single-membrane bound organelles of eukaryotic cells that are critically involved in the digestion of macromolecules mainly delivered by endocytosis and autophagy (1). Degradation occurs through hydrolytic enzymes requiring a low-pH environment (~4.5–5.0). Lysosome-associated membrane proteins play a critical role in lysosomal function (2). Lysosome-associated membrane protein 1 and 2 (LAMP1 and LAMP2) are two abundant lysosomal membrane proteins involved in regulating lysosomal motility during lysosome-phagosome fusion and cholesterol trafficking (3,4). CD63/LAMP3, a member of the tetraspanin family, is expressed on the surface of late endosomes and lysosomes and is also enriched in secreted exosomes (5). Lysosomal integral membrane protein type 2 (LIMP-2, also known as SCARB2) functions in cholesterol transport from the lysosome to the membrane, an important function as cholesterol contributes to various biophysical properties of cell membranes and may contribute to various diseases (6). Mucolipin-1 (MCOLN1, TRPML1), a member of the transient receptor potential ion channel superfamily, is a lysosomal/late endosome Ca2+ efflux channel that is important in autophagy and lysosomal biogenesis (7).

The acidic environment of the lysosome is maintained by H+ ATPase (V-ATPase), a large heteromultimeric proton pump located on the lysosomal transmembrane. Vacuolar ATPase enzymes are large, multimeric protein complexes with component proteins found in either the V1 peripheral domain or the V0 integral domain (8). The cytoplasmic V1 domain contains a hexamer of A and B catalytic subunits and several other protein subunits required for ATPase assembly and ATP hydrolysis. The integral V0 V-ATPase domain exhibits protein translocase activity and is responsible for proton transport across the membrane. The V-ATPase subunits ATP6V0c, ATP6V0d1, ATP6V1A, ATP6V1B2, and ATP6V1D interact with the Ragulator protein complex and are essential for amino acid induced activation of mTORC1 on the surface of lysosomes (9). Additionally, ATP6V1A has been shown to interact with SARS-CoV-2 M protein and facilitate viral infection (10,11).

Cathepsins are the most abundant lysosomal proteases that can be categorized into groups of cysteine, serine, and aspartic proteases (12). Cathepsin B (CTSB), a cysteine protease, has been associated with multiple sclerosis (13), rheumatoid arthritis (14), and pancreatitis (15). Expression can correlate with poor prognosis for a variety of forms of cancer (16). Cathepsin D (CTSD) is a lysosomal aspartyl protease that plays a role in neuronal degradation and malignant transformation, particularly in breast cancer (17,18).

β-glucocerebrosidase (GCase) is a lysosomal enzyme that catalyzes the hydrolysis of glucocerebroside into free ceramide and glucose (19). GBA mutations are the most common genetic risk factor for Parkinson’s disease (20).

  1. Schröder, B.A. et al. (2010) Proteomics 10, 4053-76.
  2. Eskelinen, E.L. et al. (2003) Trends Cell Biol 13, 137-45.
  3. Huynh, K.K. et al. (2007) EMBO J 26, 313-24.
  4. Eskelinen, E.L. et al. (2004) Mol Biol Cell 15, 3132-45.
  5. Escola, J.M. et al. (1998) J Biol Chem 273, 20121-7.
  6. Heybrock, S. et al. (2019) Nat Commun 10, 3521.
  7. Cheng, X. et al. (2010) FEBS Lett 584, 2013-21.
  8. Jefferies, K.C. et al. (2008) Arch Biochem Biophys 476, 33-42.
  9. Zoncu, R. et al. (2011) Science 334, 678-83.
  10. Gordon, D.E. et al. (2020) Nature 583, 459-468.
  11. Daniloski, Z. et al. (2021) Cell 184, 92-105.e16.
  12. Yadati, T. et al. (2020) Cells 9, 1679. doi: 10.3390/cells9071679.
  13. Bever, C.T. et al. (1994) Neurology 44, 745-8.
  14. Hashimoto, Y. et al. (2001) Biochem Biophys Res Commun 283, 334-9.
  15. Halangk, W. et al. (2000) J Clin Invest 106, 773-81.
  16. Mijanović, O. et al. (2019) Cancer Lett 449, 207-214.
  17. Nomura, T. and Katunuma, N. (2005) J Med Invest 52, 1-9.
  18. Nakanishi, H. (2003) Ageing Res Rev 2, 367-81.
  19. Ho, M.W. et al. (1973) Biochem J 131, 173-6.
  20. Sidransky, E. and Lopez, G. (2012) Lancet Neurol 11, 986-98.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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