Anti-DMC1 antibody [EPR23207-130]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DMC1 antibody [EPR23207-130] (AB245217)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling DMC1 with ab245217 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on spermatocytes of rat testis. The section was incubated with ab245217 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 citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DMC1 antibody [EPR23207-130] (AB245217)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling DMC1 with ab245217 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : No staining on mouse cerebrum. The section was incubated with ab245217 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 citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-DMC1 antibody [EPR23207-130] (AB245217)

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Rat testis tissue labeling VSIG4 with ab245217 at 1/100 (6.7 μg/ml) dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DMC1 antibody [EPR23207-130] (AB245217)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling DMC1 with ab245217 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on spermatocytes of mouse testis. The section was incubated with ab245217 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 citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-DMC1 antibody [EPR23207-130] (AB245217)

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Mouse testis tissue labeling VSIG4 with ab245217 at 1/100 (6.7 μg/ml) dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

• IP

Unknown

Immunoprecipitation - Anti-DMC1 antibody [EPR23207-130] (AB245217)

DMC1 was immunoprecipitated from 0.35 mg Mouse testis lysate 10ug with ab245217 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab245217 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution. Lane 1 : Mouse testis lysate 10ug

Lane 2 : ab245217 IP in Mouse testis lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab245217 in Mouse testis lysate

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

Exposure time : 30 seconds

All lanes:

Immunoprecipitation - Anti-DMC1 antibody [EPR23207-130] (ab245217)

Predicted band size: 38 kDa,87 kDa

false

• WB

Lab

Western blot - Anti-DMC1 antibody [EPR23207-130] (AB245217)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22156371).

Negative control : Spleen (PMID : 21085687).

Exposure time : 26 seconds

All lanes:

Western blot - Anti-DMC1 antibody [EPR23207-130] (ab245217) at 1/1000 dilution

Lane 1:

Mouse testis lysate at 20 µg

Lane 2:

Mouse spleen lysate at 20 µg

Lane 3:

Rat testis lysate at 20 µg

Lane 4:

Rat spleen 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: 38 kDa

Observed band size: 38 kDa

false

• WB

Lab

Western blot - Anti-DMC1 antibody [EPR23207-130] (AB245217)

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

No cross-reactivity with mouse Rad51 is observed.

Exposure time : Lane 3 seconds; Lane 2 : 81 seconds.

All lanes:

Western blot - Anti-DMC1 antibody [EPR23207-130] (ab245217) at 1/1000 dilution

Lane 1:

His-tagged mouse DMC1 recombinant protein (aa2-340) at 10 µg

Lane 2:

His-tagged mouse Rad51 recombinant protein (aa2-339) 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

Predicted band size: 38 kDa

Observed band size: 38 kDa

false