Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (AB313583)

Immunohistochemical analysis of paraffin-embedded Mouse perfused fixed tissue labeling Transferrin with ab313583 at 1/100 (5.21 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Negative control : No staining on mouse perfused fixed skeletal muscle.The section was incubated with ab313583 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (AB313583)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Hepa1-6 (mouse hepatoma epithelial cell) cells labelling Transferrin with ab313583 at 1/50 (10.42 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in Hepa1-6 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (AB313583)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Hepa1-6 (mouse hepatoma epithelial cell) cells labelling Transferrin with ab313583 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (AB313583)

Immunohistochemical analysis of paraffin-embedded (A) Non-perfused fix tissue labeling Transferrin with ab313583 at 1/100 (5.21 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Positive staining on plasma of (A) non-perfused fixed mouse cardiac muscle, no staining on (B) perfused fixed mouse cardiac muscle.The section was incubated with ab313583 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (AB313583)

Immunohistochemical analysis of paraffin-embedded (A) Non-perfused fix tissue labeling Transferrin with ab313583 at 1/100 (5.21 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Positive staining on hepatocytes and plasma of (A) non-perfused fixed mouse liver; only positive on hepatocytes of (B) perfused fixed mouse liver.The section was incubated with ab313583 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

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Immunoprecipitation - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (AB313583)

Transferrin was immunoprecipitated from 0.35 mg Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate with ab313583 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313583 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate Lane 2 : ab313583 IP in Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313583 in Hepa1-6 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (ab313583) at 1/30 dilution

All lanes:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

false

Exposure time: 180s

• WB

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Western blot - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (AB313583)

Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-Bovine Serum Albumin antibody ((ab192603) loading control staining at 1/1000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (ab313583) at 1/1000 dilution

All lanes:

Mouse plasma 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 1/100000 dilution

Observed band size: 76 kDa

false

Exposure time: 180s

• WB

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Western blot - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (AB313583)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : skeletal muscle (PMID : 3558394). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 81 seconds

All lanes:

Western blot - Anti-Transferrin antibody [EPR28141-177] - Serum Loading Control (ab313583) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue 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 1/100000 dilution

Observed band size: 76 kDa

false

Exposure time: 81s