Anti-SMCR8 antibody [EPR26215-5]

• Flow Cyt

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Flow Cytometry - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling SMCR8 with ab303548 at 1/500 dilution (0.1ug) (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 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling SMCR8 with ab303548 at 1/50 (10.52 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing mainly cytoplasmic and weak nuclear staining in HeLa cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/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 2ug/mL dilution.

• IP

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Immunoprecipitation - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

SMCR8 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell), whole cell lysate 10 ug with ab303548 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab303548 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell), whole cell lysate 10 ug Lane 2 : ab303548 IP in HeLa whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab303548 in HeLa whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 32 seconds The bands beneath the target band in lane2 are likely to be degraded target fragments.

All lanes:

Immunoprecipitation - Anti-SMCR8 antibody [EPR26215-5] (ab303548) at 1/1000 dilution

All lanes:

HeLa (human cervical adenocarcinoma 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 1/5000 dilution

false

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling SMCR8 with ab303548 at 1/50 (10.52 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 and weak nuclear staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/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.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh) tissue labeling SMCR8 with ab303548 at 1/500 (1.052 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Low expression control : confocal image showing no staining on mouse kidney (PMID : 11997338). The nuclear counterstain was DAPI (Blue). The section was incubated with ab303548 for 60mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh) tissue labeling SMCR8 with ab303548 at 1/500 (1.052 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on mouse hippocampus. The nuclear counterstain was DAPI (Blue). The section was incubated with ab303548 for 60mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• Flow Cyt

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Flow Cytometry - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling SMCR8 with ab303548 at 1/500 dilution (0.1ug) (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 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling SMCR8 with ab303548 at 1/2000 (0.263 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Weakly cytoplasmic staining on mouse kidney (PMID : 27103069). The section was incubated with ab303548 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling SMCR8 with ab303548 at 1/2000 (0.263 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse cerebrum (PMID : 27103069). The section was incubated with ab303548 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

SMCR8 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 ug with ab303548 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab303548 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 ug Lane 2 : ab303548 IP in NIH/3T3 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab303548 in NIH/3T3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 32 seconds The bands beneath the target band in lane2 are likely to be degraded target fragments.

All lanes:

NIH/3T3 (mouse embryonic fibroblast), 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: 32s

• WB

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Western blot - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Blocking and dilution buffer and concentration : 5% NFDM/TBST. The molecular weight observed is consistent with what has been described in the literature (PMID : 27103069). The bands beneath the target band are likely to be degraded target fragments. In lanes 1, the lysate was stored at -80℃ prior to Western Blotting. In lane 2-5, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.  Exposure time : Lane 1 : 84 seconds Lane 2-5 : 3 minutes

All lanes:

Western blot - Anti-SMCR8 antibody [EPR26215-5] (ab303548) at 1/1000 dilution

Lanes 1 - 2:

HeLa (human cervical adenocarcinoma epithelial cell) at 20 µg

Lane 3:

HEK 293T (human embryonic kidney epithelial cell) at 20 µg

Lane 4:

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

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma), whole cell fresh 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

Predicted band size: 105 kDa

Observed band size: 140 kDa

false

• WB

Supplier Data

Western blot - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Blocking and dilution buffer and concentration : 5% NFDM/TBST.  The molecular weight observed is consistent with what has been described in the literature (PMID : 27103069). The bands beneath the target band are likely to be degraded target fragments. Low expression : kidney, lung (PMID : 27103069).  Exposure time Lane 1-4 : 81 seconds Lane 5-7 : 3 minutes.

All lanes:

Western blot - Anti-SMCR8 antibody [EPR26215-5] (ab303548) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Lane 3:

Human lung tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 105 kDa

Observed band size: 140 kDa

false

• WB

Supplier Data

Western blot - Anti-SMCR8 antibody [EPR26215-5] (AB303548)

Blocking and dilution buffer and concentration : 5% NFDM/TBST. The molecular weight observed is consistent with what has been described in the literature (PMID : 27103069). The bands beneath the target band are likely to be degraded target fragments. Low expression : kidney (PMID : 27103069) In lanes 1-4, the lysates were stored at -80℃ prior to Western Blotting. In lane 5-7, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation. Exposure time : Lane 1-4 : 81 seconds Lane 5-7 : 3 minutes

All lanes:

Western blot - Anti-SMCR8 antibody [EPR26215-5] (ab303548) at 1/1000 dilution

Lane 1:

Mouse cerebral cortex tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

Rat cerebral cortex tissue lysate at 20 µg

Lane 4:

Rat kidney tissue lysate at 20 µg

Lane 5:

Mouse brain fresh tissue lysate at 20 µg

Lane 6:

Mouse liver fresh tissue lysate at 20 µg

Lane 7:

Mouse testis fresh 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

Predicted band size: 105 kDa

Observed band size: 140 kDa

false