Anti-Rfx Cas13d (N-term) antibody [EPR23965-31]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Rfx Cas13d (N-term) with ab314741 at 1/100 (4.8 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human liver. The section was incubated with ab314741 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Rfx Cas13d (N-term) with ab314741 at 1/100 (4.8 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse liver. The section was incubated with ab314741 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Rfx Cas13d (N-term) with ab314741 at 1/100 (4.8 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat liver. The section was incubated with ab314741 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T transfected with a Ruminococcus flavefaciens cas13d expression vector containing a HA-myc-tag (N-term) and a His-tag® (C-term) (Middle) / 293T cells transfected with empty expression vector containing a myc-His-tag® (Right) cells labelling Rfx Cas13d (N-term) with ab314741 at 1/500 dilution (0.1 ug)/Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells are co-stained with Myc conjugated to Alexa Fluor®647.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T transfected with a Ruminococcus flavefaciens cas13d expression vector containing a myc-His-tag® (C-term) (Middle) / 293T cells transfected with empty expression vector containing a myc-His-tag® (Right) cells labelling Rfx Cas13d (N-term) with ab314741 at 1/500 dilution (0.1 ug)/Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells are co-stained with Myc conjugated to Alexa Fluor®647.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a Ruminococcus flavefaciens cas13d expression vector containing a His tag. (B) HEK-293T transfected with empty vector containing a His tag. tissue labeling Rfx Cas13d (N-term) with ab314741 at 1/2000 (0.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a Ruminococcus flavefaciens cas13d expression vector containing a His tag, negative staining on (B) HEK-293T transfected with empty vector containing a His tag. The section was incubated with ab314741 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (Human embryonic kidney epithelial cell) transfected with a Ruminococcus flavefaciens cas13d expression vector containing a HA-myc-tag(N-term) and a His-tag®(C-term) cells labelling Rfx Cas13d (N-term) with ab314741 at 1/500 (0.96 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 positive staining in 293T transfected with a Ruminococcus flavefaciens cas13d expression vector containing a HA-myc-tag(N-term) and a His-tag®(C-term).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain at 1/100 (0.38ug/ml) dilution (Magenta). 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.

• IP

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Immunoprecipitation - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Rfx Cas13d (N-term) was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) cells transfected with a Ruminococcus flavefaciens cas13d expression vector containing a myc-His-tag®, whole cell lysate with ab314741 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314741 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (ab314741) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with a Ruminococcus flavefaciens cas13d expression vector containing a myc-His-tag®, whole cell lysate

Lane 2:

ab314741 at 1/30 IP in 293T (human embryonic kidney epithelial cell) cells transfected with a Ruminococcus flavefaciens cas13d expression vector containing a myc-His-tag®, whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab314741 in 293T cells transfected with a Ruminococcus flavefaciens cas13d expression vector containing a myc-His-tag®, 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 1/5000 dilution

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Rfx Cas13d may cleave to two fragments close to C-term, and this antibody can recognize the full length and N-terminal fragment. His-tag antibody can recognize full length and C-terminal fragment because the His-tag of this overexpression vector located on C-term.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (ab314741) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a Ruminococcus flavefaciens cas13d expression vector containing a myc-His-tag® (C-term), whole cell lysate at 10 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: 110 kDa,70 kDa,124 kDa,40 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (AB314741)

Rfx Cas13d may cleave to two fragments close to C-term, and this antibody can recognize the full length and N-terminal fragment. HA tag antibody can recognize the full length and N-terminal fragment. His-tag antibody can recognize full length and C-terminal fragment because the His-tag of this overexpression vector located on C-term, HA-myc-tag of this overexpression vector located on N-term.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-HA tag antibody [EPR22819-101] - (ab236632) staining at 1/1000 dilution.

In Western blot, Anti-6XHis tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (ab314741) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a Ruminococcus flavefaciens cas13d expression vector containing a HA-myc-tag (N-term) and a His-tag® (C-term), whole cell lysate at 10 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: 110 kDa,70 kDa,36 kDa,40 kDa

false

Exposure time: 6s