Purified naïve CD8+ T cells from Stat1+/+ or Stat1−/− mice were cultured with IL-7 (1 ng/ml) and IFN-β (10 ng/ml) for 2 to 3 days. After the culture, 2 × 105 cells were washed twice with ice-cold PBS and placed on a poly-l -lysine–coated glass slide (Sigma-Aldrich) for 10 min and allowed to adhere to the slide for 5 min at room temperature. The cells were fixed for 20 min with cold 4% paraformaldehyde in PBS, permeabilized for 5 min with 0.1% Triton X-100 in PBS, and then blocked for 15 min with 5% normal goat serum in PBS containing 1% bovine serum albumin. Cells were stained with rabbit anti-mouse mTOR (7C10; Cell Signaling Technology), rabbit anti-mouse LC3B (E7X4S; Cell Signaling Technology), rat anti-mouse LAMP2 (GL2A7; Abcam) for 45 min, washed twice, blocked, and then reincubated with anti-rabbit Alexa or anti-rat Alexa for 30 min. The final slides were washed with PBS and mounted in ProLong Gold Antifade Reagent (Invitrogen) and analyzed using a Zeiss LSM 700 laser scanning confocal microscope (Carl Zeiss) for acquiring fluorescence images.
Rat anti mouse lamp2 gl2a7
Manufactured by Abcam
Rat anti-mouse LAMP2 (GL2A7;) is a primary antibody that recognizes the mouse lysosome-associated membrane protein 2 (LAMP2). LAMP2 is a transmembrane glycoprotein that plays a role in the fusion of lysosomes with autophagosomes.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti γ adaptin, by BD (2 mentions)
Alexa fluor 640, by Jackson ImmunoResearch (2 mentions)
Mouse anti γ tubulin gtu 88, by Merck Group (2 mentions)
Alexa fluor 680, by Jackson ImmunoResearch (2 mentions)
Alexa fluor 790, by Jackson ImmunoResearch (2 mentions)
Mouse anti gfp clones 7 1 and 13, by Roche (2 mentions)
Alexa fluor 488, by Jackson ImmunoResearch (2 mentions)
Alexa fluor 594, by Jackson ImmunoResearch (2 mentions)
Rat anti ha 3f10, by Roche (2 mentions)
Ta99 mel 5, by American Type Culture Collection (2 mentions)
Poly l lysine coated glass slide, by Merck Group (1 mentions)
Lsm 700 confocal laser scanning microscope, by Zeiss (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Ab201227, by Abcam (1 mentions)
Pabg1, by Proteintech (1 mentions)
3 protocols using rat anti mouse lamp2 gl2a7
1
Immunofluorescence Analysis of mTOR, LC3B, and LAMP2 in Stat1-Deficient CD8+ T Cells
2
Characterization of Intracellular Trafficking Proteins
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Primary monoclonal antibodies used and their sources (indicated in parentheses) were: mouse anti-TYRP1 (TA99/Mel-5; American Type Culture Collection); mouse anti-γ-Tubulin (GTU-88; Sigma-Aldrich); mouse anti-γ-adaptin (610385; BD); mouse anti-GFP (clones 7.1 and 13.1; Roche); rabbit anti-AP3M1 (ab201227; Abcam); rat anti–mouse LAMP2 (GL2A7; Abcam); rat anti-mouse TfR (CD71; BD 553264); and rat anti-HA (3F10; Roche 11867423001). Primary polyclonal antibodies used and their sources were: rabbit anti-STX13 (a kind gift of Rytis Prekeris, University of Colorado, Denver, CO; Prekeris et al., 1998 (link)); rabbit anti-pallidin (a kind gift of Juan Bonifacino, National Institute of Child Health and Human Development, Bethesda, MD; Moriyama and Bonifacino, 2002 (link)); rabbit anti-PI4KIIIβ (13247-1-AP; Proteintech); rabbit anti-PI4KIIα and anti-PI4KIIβ (kind gifts of Pietro De Camilli, Yale University, New Haven, CT; Guo et al., 2003 (link)); and rabbit anti-GFP (PABG1; Chromotek); Species- and/or mouse isotype–specific secondary antibodies from donkey or goat conjugated to Alexa Fluor 488, Alexa Fluor 594, or Alexa Fluor 640 used for IFM or to Alexa Fluor 680 or Alexa Fluor 790 for immunoblots were obtained from Jackson ImmunoResearch Laboratories.
3
Antibody Panel for Cellular Protein Localization
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Primary monoclonal antibodies used and their sources (indicated in parentheses) were: mouse anti-TYRP1 (TA99/Mel-5, American Type Culture Collection; Rockville, MD); mouse anti-γ-Tubulin (GTU-88, Sigma); mouse anti-γ-adaptin (BD 610385); mouse anti-GFP (clones 7.1 and 13.1, Roche); rabbit anti-AP3M1 (ab201227; Abcam); rat anti-mouse LAMP2 (GL2A7, Abcam); rat anti-mouse TfR (CD71, BD 553264); and rat anti-HA (3F10, Roche 11867423001). Primary polyclonal antibodies used and their sources were: rabbit anti-STX13 (a kind gift of Rytis Prekeris, Univ. of Colorado, Denver, CO, USA) (Prekeris et al., 1998) ; rabbit anti-pallidin (a kind gift of Juan Bonifacino, National Institute of Child Health and Human Development, Bethesda, MD, USA) (Moriyama and Bonifacino, 2002) ; rabbit anti-PI4KIIIβ (13247-1-AP, Proteintech); rabbit anti-PI4KIIα and anti-PI4KIIβ (kind gifts of Pietro De Camilli, Yale Univ., New Haven, CT, USA) (Guo et al., 2003) ; and rabbit anti-GFP (PABG1, Chromotek); Species-and/or mouse isotype-specific secondary antibodies from donkey or goat conjugated to Alexa Fluor 488, Alexa Fluor 594, or Alexa Fluor 640 used for IFM or to Alexa Fluor 680 or Alexa Fluor 790 for immunoblots were obtained from Jackson ImmunoResearch Laboratories.
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