重组Anti-SEC23B抗体[EPR22723-24]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SEC23B antibody [EPR22723-24] (AB245212)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling SEC23B with 245212 at 1/1/1000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse liver is observed (PMID : 22745161).
The section was incubated with ab245212 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SEC23B antibody [EPR22723-24] (AB245212)

ab245212 staining SEC23B in Raw264.7 cells, with negative expression in cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab245212 at 1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SEC23B antibody [EPR22723-24] (AB245212)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MEF cells labeling SEC23B with ab245212 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocol image showing mainly cytoplasmic staining in MEFs. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SEC23B antibody [EPR22723-24] (AB245212)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MEF (Mouse embryonic fibroblast (immortalized)) cells labeling SEC23B with ab245212 at 1/500 dilution (0.1μg)/ (Red) compared with a Rabbit monoclonal IgG (ab172730) / (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IP

Unknown

Immunoprecipitation - Anti-SEC23B antibody [EPR22723-24] (AB245212)

SEC23B was immunoprecipitated from 0.35 mg MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate 20 ug with ab245212 at 1/30. Western blot was performed on the immunoprecipitate using ab245212 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.

Lane 1 : MEF whole cell lysate 20 ug.

Lane 2 : ab245212 IP in MEF whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab245212 in MEF whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-SEC23B antibody [EPR22723-24] (ab245212)

Predicted band size: 86 kDa

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• WB

Lab

Western blot - Anti-SEC23B antibody [EPR22723-24] (AB245212)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 22745161).

Exposure times : Lane 1 : 26 seconds; Lane 2 : 3 seconds; Lanes 3-4 : 6 seconds.

All lanes:

Western blot - Anti-SEC23B antibody [EPR22723-24] (ab245212) at 1/1000 dilution

Lane 1:

MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 86 kDa

false

• WB

Lab

Western blot - Anti-SEC23B antibody [EPR22723-24] (AB245212)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The lysates were kindly provided by an anonymous collaborator. Exposure time : 59 seconds.

All lanes:

Western blot - Anti-SEC23B antibody [EPR22723-24] (ab245212) at 1/1000 dilution

Lane 1:

MEF (mouse embryonic fibroblast (immortalized)) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

MEF (mouse embryonic fibroblast (immortalized)) transfected with siRNA specifically targeting SEC23B whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 86 kDa

false

• WB

Lab

Western blot - Anti-SEC23B antibody [EPR22723-24] (AB245212)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-SEC23B antibody [EPR22723-24] (ab245212) at 1/5000 dilution

Lane 1:

Purified LIF/His-tagged SEC23B recombinant protein, 10 ng

Lane 2:

His-tagged SEC23A recombinant protein, 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 86 kDa

false