Anti-TMEM16A antibody [SP31] - BSA and Azide free

• IHC

Lab

Immunohistochemistry - Anti-TMEM16A antibody [SP31] - BSA and Azide free (AB198412)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab64085).

Immunohistochemical analysis of formalin fixed paraffin embedded human GIST labelling TMEM16A with ab64085 at a concentration of 1.14 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32 mins.

ab64085 anti-TMEM16A [SP31] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM16A antibody [SP31] - BSA and Azide free (AB198412)

TMEM16A detected in paraffin-embedded sections of human submucosal glands from CF patients and control samples.

TMEM16A expression was modest in non-CF submucosal glands of non-CF samples (b) but markedly increased in tissues from CF patients (d), with a particularly strong signal (f) in histologically altered glands (e). Magnification : X630 in a, X400 in b-f. Images 4Aa and 4Ba,c,e show hematoxylin and eosin staining.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab64085)

This image is taken from Upregulation of TMEM16A Protein in Bronchial Epithelial Cells by Bacterial Pyocyanin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM16A antibody [SP31] - BSA and Azide free (AB198412)

Immunohistochemical analysis of Human Gastrointestinal Stromal Tumor tissue labelling TMEM16A with ab64085.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab64085)

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TMEM16A antibody [SP31] - BSA and Azide free (AB198412)

Intracellular Flow Cytometry analysis of PC-3 (Human prostate adenocarcinoma epithelial cell) cells labeling TMEM16A with purified ab64085 at 1/20 dilution (11.3μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab64085).

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM16A antibody [SP31] - BSA and Azide free (AB198412)

ab64085 showing expression of TMEM16A in human surface epithelium tissue. TMEM16A staining was mostly absent (b), and sometimes scanty (c) in the respiratory epithelium lining bronchi or bronchioles from CF patients (c, arrows). A weak expression was also detectable in microvessels of peri-bronchial connective tissue (b, arrows). Magnification : X400.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab64085)

This image is taken from Upregulation of TMEM16A Protein in Bronchial Epithelial Cells by Bacterial Pyocyanin.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TMEM16A antibody [SP31] - BSA and Azide free (AB198412)

This data was developed using ab64085, the same antibody clone in a different buffer formulation.

ab64085 staining TMEM16A in PC-3 (human prostate adenocarcinoma epithelial cell) cells. The cells were fixed with 4% formaldehyde, permeabilized in 100% methanol. The cells were then incubated with ab64085 at 1/20 dilution, followed by secondary antibody ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used for counterstain at 1/200 dilution (Red). Nuclear DNA was labelled in blue with DAPI.

Confocal image showing membranous and cytoplasmic staining in PC-3 cell line.

Low expression control : LnCaP(PMID : 29899325)

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• WB

Lab

Western blot - Anti-TMEM16A antibody [SP31] - BSA and Azide free (AB198412)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lane 1 : 20 seconds Lane 2 : 10 seconds The observed molecular weights are consistent with PMID : 22685202. This data was developed using ab64085, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-TMEM16A antibody [SP31] (<a href='/en-us/products/primary-antibodies/tmem16a-antibody-sp31-ab64085'>ab64085</a>) at 1/1000 dilution

Lane 1:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Capan-1 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 114 kDa

Observed band size: 130 kDa,260 kDa

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