Anti-HLA Class 1 ABCG antibody [EPR29051-62]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling HLA Class 1 ABCG with ab323616 at 1/4000 (0.122 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of human cerebrum (PMID : 36437414).
The section was incubated with ab323616 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Immunohistochemical analysis of 10% PFA fixed frozen Human spleen (fresh frozen) tissue labeling HLA Class 1 ABCG with ab323616 followed by ready to use LeicaDS9800 (Bond™ PolymerRefine Detection).

Positive staining in human spleen. The section was fixed in 10% PFA and incubated with ab323616 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) was used.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Immunohistochemical analysis of 10% PFA fixed frozen Human lung (fresh frozen) tissue labeling HLA Class 1 ABCG with ab323616 followed by ready to use LeicaDS9800 (Bond™ PolymerRefine Detection).

Positive staining in human lung. The section was fixed in 10% PFA and incubated with ab323616 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) was used.

• Flow Cyt

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Flow Cytometry - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Flow cytometric analysis of A431 (human epidermoid carcinoma epithelial cell) (Right) / K562 (chronic myelogenous leukemia) (Left) cells labelling HLA Class 1 ABCG with ab323616 at 1/50 dilution (1ug) (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).

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

Gated on viable cells.
Low expression : K562 (PMID : 25960936).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling HLA Class 1 ABCG with ab323616 at 1/4000 (0.122 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.
The section was incubated with ab323616 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Immunohistochemical analysis of paraffin-embedded Human adenocarcinoma of colon tissue labeling HLA Class 1 ABCG with ab323616 at 1/4000 (0.122 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human adenocarcinoma of colon (PMID : 31113192).
The section was incubated with ab323616 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Immunohistochemical analysis of paraffin-embedded (A) A431 (human epidermoid carcinoma epithelial cell) cell pellet, (B) T-47D (human ductal breast epithelial tumor epithelial cell) cell pellet, (C) K-562 (human chronic myelogenous leukemia lymphoblast) cell pellet and (D) JAR (human placenta choriocarcinoma epithelial cell) cell pellet labeling HLA Class 1 ABCG with ab323616 at 1/2000 (0.244 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) A431 cell pellet and (B) T-47D cell pellet, no staining on (C) K-562 cell pellet and (D) JAR cell pellet.
The section was incubated with ab323616 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Immunohistochemical analysis of paraffin-embedded Human endometrioid carcinoma tissue labeling HLA Class 1 ABCG with ab323616 at 1/4000 (0.122 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human endometrioid carcinoma.
The section was incubated with ab323616 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling HLA Class 1 ABCG with ab323616 at 1/4000 (0.122 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen.
The section was incubated with ab323616 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling HLA Class 1 ABCG with ab323616 at 1/4000 (0.122 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of human lung (PMID : 36437414).
The section was incubated with ab323616 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Supplier Data

Flow Cytometry - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Flow cytometric analysis of T-47D (human ductal breast epithelial tumor epithelial cell) (Right) / JAR (human placenta choriocarcinoma epithelial cell) (Left) cells labeling HLA Class 1 ABCG with ab323616 at 1/50 dilution (1ug) (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).

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

Gated on viable cells.
Low expression : JAR.

• WB

Supplier Data

Western blot - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

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

Low expression : K-562 (PMID : 25960936), IMR-32, JAR, MCF7.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 24173243).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

Exposure time : Lanes 1-2 : 1 second, lanes 3-7 : 3 seconds

All lanes:

Western blot - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (ab323616) at 1/1000 dilution

Lane 1:

KARPAS-299 (human T cell lymphoma cell) whole cell lysate at 20 µg

Lane 2:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 4:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 5:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 6:

JAR (human placenta choriocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell 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

Observed band size: 40 kDa,41 kDa,15 kDa

false

• WB

Supplier Data

Western blot - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

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 : 24173243).

Exposure time : Lanes 1-2 : 3 seconds, lanes 3-5 : 37 seconds

All lanes:

Western blot - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (ab323616) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human lymph node tissue lysate at 20 µg

Lane 3:

Human testis tissue lysate at 20 µg

Lane 4:

Human bladder tissue lysate at 20 µg

Lane 5:

Human cerebellum tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 40 kDa,41 kDa

false

• Dot

Supplier Data

Dot Blot - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (AB323616)

Dot blot analysis of HLA Class 1 ABCG using ab323616 at 1/1000 (0.488 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.

Lane 1 : LIF-tagged human HLA Class 1 A fragment
Lane 2 : LIF-tagged human HLA Class 1 B fragment
Lane 3 : LIF-tagged human HLA Class 1 C fragment
Lane 4 : LIF/His-tagged human HLA Class 1 E fragment
Lane 5 : His-tagged human HLA Class 1 F fragment
Lane 6 : LIF-tagged human HLA Class 1 G fragment

Exposure time : 180 seconds.

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

This antibody reacts with human HLA Class 1 A, HLA Class 1 B, HLA Class 1 C and HLA Class 1 G but not HLA Class 1 E and HLA Class 1 F.

All lanes:

Dot Blot - Anti-HLA Class 1 ABCG antibody [EPR29051-62] (ab323616) at 1/1000 dilution

Lane 1:

LIF-tagged human HLA Class 1 A fragment

Lane 2:

LIF-tagged human HLA Class 1 B fragment

Lane 3:

LIF-tagged human HLA Class 1 C fragment

Lane 4:

LIF/His-tagged human HLA Class 1 E fragment

Lane 5:

His-tagged human HLA Class 1 F fragment

Lane 6:

LIF-tagged human HLA Class 1 G fragment

false

Exposure time: 180s