- 11-1078-41 - Invitrogen Antibodies LAMP2 antibody

Submitted references Improved Autophagic Flux in Escapers from Doxorubicin-Induced Senescence/Polyploidy of Breast Cancer Cells.

Bojko A, Staniak K, Czarnecka-Herok J, Sunderland P, Dudkowska M, Śliwińska MA, Salmina K, Sikora E
International journal of molecular sciences 2020 Aug 24;21(17)

Lysosomal membrane permeabilization is involved in oxidative stress-induced apoptotic cell death in LAMP2-deficient iPSCs-derived cerebral cortical neurons.

Law CY, Siu CW, Fan K, Lai WH, Au KW, Lau YM, Wong LY, Ho JCY, Lee YK, Tse HF, Ng KM
Biochemistry and biophysics reports 2016 Mar;5:335-345

Detection of T-cell degranulation: CD107a and b.

Betts MR, Koup RA
Methods in cell biology 2004;75:497-512

LAMP-1 and LAMP-2, but not LAMP-3, are reliable markers for activation-induced secretion of human mast cells.

Grützkau A, Smorodchenko A, Lippert U, Kirchhof L, Artuc M, Henz BM
Cytometry. Part A : the journal of the International Society for Analytical Cytology 2004 Sep;61(1):62-8

Primary LAMP-2 deficiency causes X-linked vacuolar cardiomyopathy and myopathy (Danon disease).

Nishino I, Fu J, Tanji K, Yamada T, Shimojo S, Koori T, Mora M, Riggs JE, Oh SJ, Koga Y, Sue CM, Yamamoto A, Murakami N, Shanske S, Byrne E, Bonilla E, Nonaka I, DiMauro S, Hirano M
Nature 2000 Aug 24;406(6798):906-10

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of CD107b (LAMP-2) was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CD107b(LAMP-2), FITC, Mouse Monoclonal antibody (Product # 11-1078-41) at 0.25 µg/mL in 0.1% BSA and incubated at 4 degree Celsius overnight (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents the isotype control. The images were captured at 60X magnification.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of CD107b (LAMP2) was achieved by transfecting HeLa cells with CD107b (LAMP2) specific siRNAs (Silencer® select Product # s533069, s8086 ). Immunofluorescence analysis was performed using untransfected HeLa cells (panels a, d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with CD107b (LAMP2) specific siRNAs (panel c,f). Cells were fixed, permeabilized, and probed with CD107b (LAMP-2) Monoclonal Antibody (eBioH4B4 (H4B4)), FITC, eBioscience™ (Product # 11-1078-41, 1:250 dilution). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962) and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Reduction of specific cytoplasmic localization was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to CD107b (LAMP2). The images were captured at 60X magnification.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Intracellular staining of the Jurkat cell line with Mouse IgG1 K Isotype Control FITC (Product # 11-4714-42) (open histogram) or Anti-Human CD107b (LAMP-2) FITC (filled histogram). Total cells were used for analysis.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Autophagic flux resumption in descendants of polyploid/senescent MDA-MB-231 and MCF-7 cells. Cells were treated with 100 nM doxorubicin for one day, then cultured in a fresh medium and analyzed on subsequent days. ( a , c ) Representative western blots showing autophagy protein levels of p-ULK1 (S757), ULK1, LC3B, LAMP-2 and SQSTM1/p62 in MDA-MB-231 cells ( a ) and MCF-7 cells ( c ). ( b , d ) Quantitative analysis of autophagic index based on LC3B protein levels in untreated and bafilomycin A- or chloroquine-treated MDA-MB-231 cells ( b ) and MCF-7 cells ( d ) with representative western blots showing LC3B protein levels. Bars: mean value, error bars: SEM, n = 4. Statistical significance (in relation to control): $ p < 0.051, * p < 0.05, ** p < 0.01. ( e ) Transient accumulation of autophagic vesicles. TEM images show typical MDA-MB-231cells on the subsequent days following treatment (upper panel) and their magnified parts with autophagic vesicles (red arrows) and lipofuscin particles (red asterisks).
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 LAMP2-deficient neurons showed increase levels of cytosolic cathepsin L. (A) Western blot analysis revealed the elevated level of active cathepsin L in the LAMP2-deficient iPSCs-derived cortical neurons under non-stressed condition. (B) Western blot analysis showed that oxidative stress increased the abundance of active cathepsin L in the cytosols. N =3; *: P
Conjugate: Green dye

11-1078-41

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