Cd107a 1d4b

Manufactured by BD

CD107a (1D4B) is a monoclonal antibody that binds to the cell surface protein CD107a, also known as LAMP-1. CD107a is a lysosome-associated membrane protein that is expressed on the surface of activated T cells and natural killer (NK) cells. The function of this antibody is to detect and quantify CD107a-positive cells, which can be used as a marker of cell activation and degranulation.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Anti-CD107a-FITC antibody (1D4B, (2 citations) Anti-CD107a (1D4B) (2 citations) Anti-CD107a/b antibodies (1D4B, (2 citations) CD107a FITC (clone 1D4B; (2 citations)

8 protocols using cd107a 1d4b

Isolation and Characterization of Uterine Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell suspensions of uterine tissues were prepared using either mechanical dissociation followed by centrifugation through a Lympholyte M (Cedarlane) gradient according to the manufacturers recommendations or by enzymatic digestion using Liberase TM (Roche) as described previously (Collins et al., 2009 (link)). Cells were typed using fluorochrome-conjugated antibodies with specificity for CD45 (30-F11), NK1.1 (PK138), CD11b (M1/70), CD27 (LG.3A10), CD49a (HMα1), CD49b (DX5), KLRG1 (MAFA), Ly6C (HK1.4), Gr-1 (RB6-8C5), I-A/I-E (M5/114.15.2), F4/80 (BM8), CD11c (N418), CD3ε (145-2C11), CD4 (GK1.5), CD8a (53-6.7), CD19 (1D3), CD122 (5H4), NKp46 (29A1.4), IL-6 (MP5-20F3), CD107a (1D4B), and Ki-67 (B56) purchased from BD PharMingen, BioLegend, or eBioscience. For intracellular and intranuclear antigens, Fix & Perm and Foxp3 staining buffer set (both eBioscience) were used, respectively. For estimation of total cell number and leukocyte composition, enzymatic digestion was preferred as it yielded higher amounts of non-lymphocytes and did not exclude cells based on size or density. For phenotyping of NK cells, mechanical dissociation followed by Lympholyte M was used in order to prevent degradation of surface proteins.

Kieckbusch J., Balmas E., Hawkes D.A, & Colucci F. (2015). Disrupted PI3K p110δ Signaling Dysregulates Maternal Immune Cells and Increases Fetal Mortality In Mice. Cell Reports, 13(12), 2817-2828.

+ Open protocol

+ Expand

Multiparameter Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Alive/dead stain was included to measure viability (Zombie Yellow, BioLegend). Cells were blocked with anti-FcγRIII (2.4G2) prior to surface staining. Cells were fixed with 1% paraformaldehyde for surface analysis or fixed with Cytofix/Cytoperm (BD Bioscience) for intracellular staining. For intranuclear staining, cells were fixed in Cytofix/Cytoperm and Cytoperm Plus buffer (BD) and DNase treated. For phospho mTOR staining, cells were fixed with 2% paraformaldehyde and permeabilized with methanol. For phospho AMPK staining, cells were fixed with Cytofix/Cytoperm (BD Bioscience), and stained overnight at 4C with anti-pAMPK. Data was acquired on a Cytek-modified FACScan (BD and Cytek) or LSRFortessa (BD); data was analyzed using FlowJo 7.6.5. Geometric mean fluorescence intensity and % proliferated were calculated using FlowJo proliferation analysis (see below). The following fluorochrome-conjugated antibodies were used: BrdU (BrdU kit, BD), CD107a (1D4B, BD), CD3ε (145-2C11, BD/BioLegend), CD11b (M1/70, BD), CD226/DNAM-1 (10E5, BioLegend), CD27 (LG.3A10, BD), CD45.1 (A20 Biolegend), anti-human IFN-γ (XMG1.2, BioLegend), Ly49C/I (5E6, BD), Ly49H (3D10, BD/BioLegend), NK1.1 (PK136, BD), NKG2A/C/E (20d5, BD), NKG2D (CX5, Biolegend), NKp46 (29A1.4, BD), pAMPKa, (40H9, Cell Signaling), and pMTOR (MRRBY, Invitrogen).

Mah-Som A.Y., Keppel M.P., Tobin J.M., Kolicheski A., Saucier N., Sexl V., French A.R., Wagner J.A., Fehniger T.A, & Cooper M.A. (2021). Reliance on Cox10 and oxidative metabolism for antigen-specific NK cell expansion. Cell reports, 35(9), 109209.

+ Open protocol

+ Expand

Multiparameter Flow Cytometry of Tumor-Infiltrating Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorochrome-conjugated anti-PD-L1 (10 F.9 G2), anti-CD45 (30-F11), anti-CD11b (M1/70), anti-CD8 (53-6.7), anti-CD4 (GK1.5), anti-CD3 (145-2C11), anti-Tim3 (RMT3-23) and anti-TNFα (MP6-XT22) were from Biolegend. Anti-PD1 (J43) was from eBioscience. CD107 a (1D4B) and anti-IFNγ (XMG1.2) was from BD Bioscience. For selection of tumor cells, after Percoll performed to isolate tumor-infiltrating lymphocytes, cells from the upper phase were collected. First, tumor cells were gated based on FSC/SSC parameters using in vitro tumor cell line as reference. Then CD45-7AAD- viable tumor cells were selected. All events were acquired by a BD LSR-II cytometer equipped with BD FACSDiva software (BD Biosciences) and data were analyzed using FlowJo software (Tree Star, Ashland, Oregon).

Dosset M., Vargas T.R., Lagrange A., Boidot R., Végran F., Roussey A., Chalmin F., Dondaine L., Paul C., Lauret Marie-Joseph E., Martin F., Ryffel B., Borg C., Adotévi O., Ghiringhelli F, & Apetoh L. (2018). PD-1/PD-L1 pathway: an adaptive immune resistance mechanism to immunogenic chemotherapy in colorectal cancer. Oncoimmunology, 7(6), e1433981.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Flow cytometry data were acquired on a FACSCanto (BD Biosciences, San Diego, CA) and analyzed with FlowJo software (Tree Star, Ashland, OR). To determine expression of cell surface proteins, mAb were incubated at 4°C for 20–30 min and cells were fixed using Cytofix/Cytoperm Solution (BD Biosciences) and, in some instances followed by mAb incubation to detect intracellular proteins. The following mAb clones were used: NK1.1 (PK136, eBioscience), CD3 (17A2, eBioscience), Ly49H (3D10, eBioscience), Ly49D (4E5, eBioscience), NKp46 (29A1.4, eBioscience), CD27 (LG.7F9, eBioscience), CD11b (M1/70, eBioscience), IFN-γ (XMG1.2; eBioscience), Granzyme B (MHGB04, Invitrogen), CD107a (1D4B, BD Pharmingen), AKT1 (55/PKBa/AKT, BD Pharmigen), pS473 (M89-61, BD Pharmigen).
Intracellular cytokine staining: For direct ex vivo, staining cells were incubated for 1 additional hour in the presence of Brefeldin A (BFA) before surface and intracellular IFN-γ staining. For cytokine staining following in vitro stimulation BFA was added during the last hour of stimulation. Intracellular signaling staining: For detection of intracellular AKT and pAKT (pS473) cells were methanol fixed and permeabilized according to BD protocol. Apoptosis was evaluated using Vybrant FAM Caspase-3/7 Assay Kit (Invitrogen) according to manufacturer’s protocol.

Jensen I.J., Winborn C.S., Fosdick M.G., Shao P., Tremblay M.M., Shan Q., Tripathy S.K., Snyder C.M., Xue H.H., Griffith T.S., Houtman J.C, & Badovinac V.P. (2018). Polymicrobial sepsis influences NK-cell-mediated immunity by diminishing NK-cell-intrinsic receptor-mediated effector responses to viral ligands or infections. PLoS Pathogens, 14(10), e1007405.

+ Open protocol

+ Expand

Murine Splenocyte Isolation and Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols

For flow cytometry, cells were isolated from mouse spleens and mechanically dissociated with a 40-µm cell strainer (Greiner Bio-One). Red blood cells were lysed with RBC lysing buffer (Sigma). Cells were plated in 96-well plates (Corning) (1×10⁶ cells/well) with brefeldin A (Biolegend), monensin (Biolegend), CD107a (1D4B), and CD107b (ABL-93; both BD Biosciences at 0.1 μg/well) in the presence or absence of a pool of HER2 peptides for 6 hours as previously described (29 ). Cells were stained with Fixable viability dye (Invitrogen) and additional fluorochrome-conjugated antibodies. For intracellular staining, a FoxP3 Fix/Perm kit was used according to the manufacturer’s instructions (eBioscience). Antibodies used include: CD45 (30F11), CD8β (YTS156.7.7), CD4 (RM4–5), CD44 (IM7), FoxP3 (FJK-16S), IFNγ (XMG1.2), and granzyme B (GB11) (all Biolegend). Data were collected using an LSR II flow cytometer (BD Bioscience) and analyzed with FlowJo software (Tree Star).

Crosby E.J., Gwin W., Blackwell K., Marcom P.K., Chang S., Maecker H.T., Broadwater G., Hyslop T., Kim S., Rogatko A., Lubkov V., Snyder J.C., Osada T., Hobeika A.C., Morse M.A., Lyerly H.K, & Hartman Z.C. (2019). Vaccine-induced memory CD8+ T cells provide clinical benefit in HER2 expressing breast cancer: a mouse to human translational study. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(9), 2725-2736.

+ Open protocol

+ Expand

Mouse NK Cell Phenotyping and Functional Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

All surface antigen staining procedures were performed for 20 min at 4°C in FACS buffer (PBS + 2% FBS). Data were acquired from an LSR Fortessa flow cytometer (BD Biosciences) and analysed using FlowJo v9.9.6 (Tree Star). The antibodies used for surface staining include CD3 (145-2C11), NKp46 (29A1.4) from BD Biosciences, and NK1.1 (PK136) from BioLegend. Antibodies used for in vitro stimulation and killing assays include NKp46 (polyclonal; R&D Systems), NK1.1 (PK136; BioLegend), CD107a (1D4B; BD Biosciences), and IFN-γ (XMG1.2; BD Biosciences). Dead cells were determined using a LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Invitrogen). Rat anti-mouse antibodies used for mouse NK cell enrichment obtained from BioLegend include CD3 (17A2), CD19 (6D5), Ter119 (TER-119), Gr-1 (RB6.8C5), CD4 (RM4–5), and CD8 (53–6.7). The secondary antibody used to cross-link mouse NK1.1 antibodies in the calcium flux experiments is goat anti-mouse IgG (H + L) polyclonal antibody (Jackson Immunoresearch).

Luu T.T., Schmied L., Nguyen N.A., Wiel C., Meinke S., Mohammad D.K., Bergö M., Alici E., Kadri N., Ganesan S, & Höglund P. (2021). Short-term IL-15 priming leaves a long-lasting signalling imprint in mouse NK cells independently of a metabolic switch. Life Science Alliance, 4(4), e202000723.

+ Open protocol

+ Expand

Cytokine Profiling in Transduced Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were incubated for 30 minutes at 4°C in PBS supplemented with 2% bovine growth serum and 0.01% sodium azide. For intracellular cytokine staining, splenocytes were re-stimulated with OVA_257–264 (InvivGen vac-sin) or GP_33–41 peptide (Anaspec) for 4 hours at 37°C with Protein Transport Inhibitor Cocktail (eBioscience) added after 1 hour of incubation. CD107a (1D4B, BD Biosciences) antibody was included in the media for the entirety of the stimulation to detect surface expression as a surrogate of degranulation. To better preserve the ametrine reporter signal in transduced populations, samples were fixed and permeabilized using the Cytofix/Cytoperm Fixation/Permeabilization kit (BD). Non-transduced populations were fixed and permeabilized using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience). Stained cells were analyzed using LSRFortessa X-20 or LSRFortessa cytometers (BD) and FlowJo software (TreeStar). Cell sorting was performed on FACSAria or FACSAria Fusion instruments (BD).

Quon S., Yu B., Russ B.E., Tsyganov K., Nguyen H., Toma C., Heeg M., Hocker J.D., Milner J.J., Crotty S., Pipkin M.E., Turner S.J, & Goldrath A.W. (2023). DNA architectural protein CTCF facilitates subset-specific chromatin interactions to limit the formation of memory CD8+ T cells. Immunity, 56(5), 959-978.e10.

+ Open protocol

+ Expand

Intracellular Cytokine Staining of CD8+ T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were incubated for 30 min at 4°C in PBS supplemented with 2% bovine growth serum and 0.1% sodium azide. Intracellular staining was performed using the BD Cytofix/Cytoperm Solution Kit (BD Biosciences). For cytokine staining, siIEL CD8⁺ T cells were incubated for 3 hours at 37°C in RPMI-1640 media containing 10% (v/v) bovine growth serum with 10 nM GP_33-41 peptide and Protein Transport Inhibitor (eBioscience) was added after 1 hour of incubation. CD107a (1D4B, BD Biosciences) antibody was included in the media for the entirety of the stimulation to detect surface expression as a surrogate of degranulation. Stained cells were analyzed using LSRFortessa or LSRFortessa X-20 cytometers (BD) and FlowJo software (TreeStar). All sorting was performed on BD FACSAria or BD FACSAria Fusion instruments.

Milner J.J., Toma C., He Z., Kurd N.S., Nguyen Q.P., McDonald B., Quezada L., Widjaja C.E., Witherden D.A., Crowl J.T., Shaw L.A., Yeo G.W., Chang J.T., Omilusik K.D, & Goldrath A.W. (2020). Heterogenous populations of tissue-resident CD8+ T cells are generated in response to infection and malignancy. Immunity, 52(5), 808-824.e7.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!