MA5-11900

antibody from Invitrogen Antibodies

Targeting: ST6GAL1 SIAT1

Provider product page for MA5-11900

Western blot

Antibody data

Antibody Data
Antigen structure
References [5]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [2]
Immunohistochemistry [1]
Flow cytometry [2]
Other assay [1]

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Product number: MA5-11900 - Provider product page
Provider: Invitrogen Antibodies
Product name: ST6GAL1 Monoclonal Antibody (LN1)
Antibody type: Monoclonal
Antigen: Other
Description: MA5-11900 targets CDw75 in FACS, IF, WB and IHC (P) applications and shows reactivity with Human and Mouse samples. Antibody does not react with rat. The molecular weight of the antigen is 53 & 87 kDa.
Antibody clone number: LN1
Concentration: 0.2 mg/mL

ST6GAL1 protein structure - MA5-11900 shown in red.

Submitted references ST6GAL1 and α2-6 Sialylation Regulates IL-6 Expression and Secretion in Chronic Obstructive Pulmonary Disease.

Krick S, Helton ES, Easter M, Bollenbecker S, Denson R, Zaharias R, Cochran P, Vang S, Harris E, Wells JM, Barnes JW
Frontiers in immunology 2021;12:693149

Identification of proteins with the CDw75 epitope in human colorectal cancer.

Mariño-Crespo Ó, Fernández-Briera A, Gil-Martín E
Oncology letters 2018 Jan;15(1):580-587

Propagation of oestrogen receptor-positive and oestrogen-responsive normal human breast cells in culture.

Fridriksdottir AJ, Kim J, Villadsen R, Klitgaard MC, Hopkinson BM, Petersen OW, Rønnov-Jessen L
Nature communications 2015 Nov 13;6:8786

Antibodies are forever: a study using 12-26-year-old expired antibodies.

Argentieri MC, Pilla D, Vanzati A, Lonardi S, Facchetti F, Doglioni C, Parravicini C, Cattoretti G
Histopathology 2013 Dec;63(6):869-76

Synthesis and expression of CDw75 antigen in human colorectal cancer.

Costa-Nogueira C, Villar-Portela S, Cuevas E, Gil-Martín E, Fernández-Briera A
BMC cancer 2009 Dec 10;9:431

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of CDw75 was performed by loading 25 µg of Hela (lane 1), Raji (lane 2) and BAF-3 (lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a CDw75 monoclonal antibody (Product # MA5-11900) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~53 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CDw75 (green) showing staining in the membrane and cytoplasm of BAF-3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CDw75 monoclonal antibody (Product # MA5-11900) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CDw75 (green) showing staining in the membrane of Ramos cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CDw75 monoclonal antibody (Product # MA5-11900) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human tonsil stained with CDw75 antibody using peroxidase-conjugate and AEC chromogen. Note cell membrane and cytoplasmic staining of B cells in germinal center.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of CDw75 in BAF-3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a CDw75 monoclonal antibody (Product # MA5-11900) at a dilution of 2 µg/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of CDw75 in PBMC cells (green) compared to an isotype control (blue). Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 µL of cell solution was added to each tube at a dilution of 2x10^7 cells/mL, followed by the addition of 50 µL of isotype control and primary antibody (Product # MA5-11900) at a dilution of 2 µg/test. Cells were incubated for 30 min at 4ºC and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated secondary antibody for 30 min at 4ºC in the dark. FACS analysis was performed using 400 µL of cell buffer.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Inhibition of ST6GAL1 secretion reduces alpha2-6 sialylation and increases IL-6 secretion similar to CSE. (A) Flow cytometric analysis of alpha2-6 sialylation using SNA-FITC labeling of HBECs (CTRL, ST6GAL1 KD, and ST6GAL1 OE) following treatment with and without iBACE and CSE for 72 hours. (B) 10,000 events were collected for each group, analyzed, and shown as the geometric mean. (C) Densitometry and a representative slot blot of secreted ST6GAL1 collected from conditioned medium following incubation with iBACE, CSE, or both. As a loading control, the supernatants were normalized to total cellular protein. (D) IL-6 secretion was determined from conditioned medium using an ELISA kit to the IL-6 ligand following same treatments as (C) . Experiments were performed in triplicate using three separate experiments. SNA, Sambucus Nigra Lectin; CTRL, control; iBACE, beta-site amyloid precursor protein cleaving enzyme 1 inhibitor; and CSE, cigarette smoke extract. All bar graphs are means +- SEM with *p < 0.05, **p < 0.01 and ***p < 0.001.