Anti-MHC Class 1 antibody [MJF-D30007-535]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunohistochemical analysis of paraffin-embedded (A) : HEK-293T (human embryonic kidney epithelial cell) cells transfected with a mouse D-B expression vector containing a his tag. (B) : HEK-293T cells transfected with a mouse D-D expression vector containing a his tag. (C) : HEK-293T cells transfected with a mouse α chain expression vector containing a his tag. (D) : HEK-293T cells transfected with a mouse D-P expression vector containing a his tag. (E) : HEK-293T cells transfected with a mouse D-K expression vector containing a his tag. (F) : HEK-293T cells transfected with empty vector containing a his tag. tissue labeling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/5000 (0.1 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T cells transfected with a mouse D-B expression vector containing a his tag, (B) HEK-293T cells transfected with a mouse D-D expression vector containing a his tag, (C) HEK-293T cells transfected with a mouse α chain expression vector containing a his tag, (D) HEK-293T cells transfected with a mouse D-P expression vector containing a his tag, and (E) HEK-293T cells transfected with a mouse D-K expression vector containing a his tag, no staining on (F) HEK-293T cells transfected with empty vector containing a his tag.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunohistochemical analysis of paraffin-embedded (A) : HEK-293T cells transfected with a mouse H2-L expression vector containing a his tag. (B) : HEK-293T cells transfected with a mouse H2-Q10 expression vector containing a his tag. (C) : HEK-293T cells transfected with a mouse H2-Q8 expression vector containing a his tag. (D) : HEK-293T cells transfected with a mouse H2-Q7 expression vector containing a his tag. (E) : HEK-293T cells transfected with a mouse PH-2D expression vector containing a his tag. tissue labeling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/5000 (0.1 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T cells transfected with a mouse H2-L expression vector containing a his tag, (B) HEK-293T cells transfected with a mouse H2-Q10 expression vector containing a his tag, (C) HEK-293T cells transfected with a mouse H2-Q8 expression vector containing a his tag, (D) HEK-293T cells transfected with a mouse H2-Q7 expression vector containing a his tag, and (E) HEK-293T cells transfected with a mouse PH-2D expression vector containing a his tag.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunohistochemical analysis of paraffin-embedded Mouse large B cell lymphoma tissue labeling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/2000 (0.251 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse large B cell lymphoma.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/2000 (0.251 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on vascular endothelium of mouse skeletal muscle.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/2000 (0.251 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunohistochemical analysis of paraffin-embedded (A) : HEK-293T cells transfected with a mouse K-B expression vector containing a his tag. (B) : HEK-293T cells transfected with a mouse K-D expression vector containing a his tag. (C) : HEK-293T cells transfected with a mouse K-K expression vector containing a his tag. (D) : HEK-293T cells transfected with a mouse K-Q expression vector containing a his tag. (E) : HEK-293T cells transfected with a mouse K-W28 expression vector containing a his tag. tissue labeling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/5000 (0.1 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T cells transfected with a mouse K-B expression vector containing a his tag, (B) HEK-293T cells transfected with a mouse K-D expression vector containing a his tag, (C) HEK-293T cells transfected with a mouse K-K expression vector containing a his tag, (D) HEK-293T cells transfected with a mouse K-Q expression vector containing a his tag, and (E) HEK-293T cells transfected with a mouse K-W28 expression vector containing a his tag.

The primary antibody was incubated 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

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized EL4 (mouse lymphoma T lymphocyte) Raw 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/500 (1.002 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in EL4 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : Raw264.7. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 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 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) transfected with an empty expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-B alpha chain expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-D alpha chain expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, alpha chain expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-P alpha chain expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-K alpha chain expression vector containing a myc-His-tag® cells labelling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/1000 (0.501 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-B alpha chain, D-D alpha chain, alpha chain, D-P alpha chain and D-P alpha chain expression vectors containing a myc-His-tag® (shown in green), showing no staining in 293T cells transfected with an empty expression vector containing a myc-His-tag®. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Panel A : 293T cells transfected with an empty expression vector containing a myc-His-tag®. Panel B : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-B alpha chain expression vector containing a myc-His-tag®. Panel C : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-D alpha chain expression vector containing a myc-His-tag®. Panel D : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, alpha chain expression vector containing a myc-His-tag®. Panel E : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-P alpha chain expression vector containing a myc-His-tag®. Panel F : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-K alpha chain expression vector containing a myc-His-tag®.

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 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 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-B alpha chain expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-D alpha chain expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-K alpha chain expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-Q alpha chain expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-W28 alpha chain expression vector containing a myc-His-tag® cells labelling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/1000 (0.501 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in 293T cells transfected with a mouse H-2 class I histocompatibility antigen, D-P alpha chain, D-K alpha chain, K-B alpha chain, K-D alpha chain, K-K alpha chain, K-Q alpha chain and K-W28 alpha chain expression vectors containing a myc-His-tag® (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Panel G : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-B alpha chain expression vector containing a myc-His-tag®. Panel H : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-D alpha chainexpression vector containing a myc-His-tag®. Panel I : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-K alpha chain expression vector containing a myc-His-tag®. Panel J : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-Q alpha chain expression vector containing a myc-His-tag® Panel K : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, K-W28 alpha chain expression vector containing a myc-His-tag®

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 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 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells transfected with a mouse H-2 class I histocompatibility antigen, L-D alpha chain (H2-L) expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, Q10 alpha chain (H2-Q10)expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, Q8 alpha chain (H2-Q8) expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, Q8 alpha chain (H2-Q7) expression vector containing a myc-His-tag® 293T cells transfected with a mouse H-2 class I histocompatibility antigen, alpha chain, Clone PH-2D-2 expression vector containing a myc-His-tag® cells labelling MHC Class 1 H2 Db/H2-D1 with ab326943 at 1/1000 (0.501 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in 293T cells transfected with a mouse H-2 class I histocompatibility antigen, L-D alpha chain (H2-L), Q10 alpha chain (H2-Q10), Q8 alpha chain (H2-Q8), Q8 alpha chain (H2-Q7) and alpha chain, Clone PH-2D-2 expression vectors containing a myc-His-tag® (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Panel L : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, L-D alpha chain (H2-L) expression vector containing a myc-His-tag® Panel M : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, Q7 alpha chain (H2-Q7) expression vector containing a myc-His-tag® Panel N : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, Q8 alpha chain (H2-Q8) expression vector containing a myc-His-tag® Panel O : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, Q7 alpha chain (H2-Q7) expression vector containing a myc-His-tag® Panel P : 293T cells transfected with a mouse H-2 class I histocompatibility antigen, alpha chain, Clone PH-2D-2 expression vector containing a myc-His-tag®

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 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 dilution.

• WB

Lab

Western blot - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

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

Low expression : Raw 264.7(PMID : 35965334)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(37-50KDa).

All lanes:

Western blot - Anti-MHC Class 1 antibody [MJF-D30007-535] (ab326943) at 1/1000 dilution

Lane 1:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

EL4 (mouse lymphoma T lymphocyte) whole cell 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:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 37-50 kDa,36 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(37-50KDa).

Exposure time : Lanes1-6 : 37 seconds

All lanes:

Western blot - Anti-MHC Class 1 antibody [MJF-D30007-535] (ab326943) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a mouse D-B expression vector containing a his tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a mouse D-D expression vector containing a his tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a mouse α chain expression vector containing a his tag, whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a mouse D-P expression vector containing a his tag, whole cell lysate at 20 µg

Lane 6:

293T cells transfected with a mouse D-K expression vector containing a his tag, whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 37-50 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(37-50KDa).

Exposure time : Lanes1-5 : 37 seconds

All lanes:

Western blot - Anti-MHC Class 1 antibody [MJF-D30007-535] (ab326943) at 1/1000 dilution

Lane 1:

293T cells transfected with a mouse K-B expression vector containing a his tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a mouse K-D expression vector containing a his tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a mouse K-K expression vector containing a his tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a mouse K-Q expression vector containing a his tag, whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a mouse K-W28 expression vector containing a his tag, whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 37-50 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-MHC Class 1 antibody [MJF-D30007-535] (AB326943)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(37-50KDa).

Exposure time : Lanes1-4 : 37 seconds, lane 5 : 8 seconds

All lanes:

Western blot - Anti-MHC Class 1 antibody [MJF-D30007-535] (ab326943) at 1/1000 dilution

Lane 1:

293T cells transfected with a mouse H2-Q10 expression vector containing a his tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a mouse H2-Q8 expression vector containing a his tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a mouse H2Q7 expression vector containing a his tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a mouse H2-L expression vector containing a his tag, whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a mouse PH-2D expression vector containing a his tag, whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 37-50 kDa,36 kDa

false