Anti-PPAR gamma antibody [EPR25862-79] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling PPAR gamma with ab310323 at 1/200 (2.575 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : no staining on human cerebrum. The section was incubated with ab310323 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 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-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human white adipose tissue labelling PPAR gamma with ab310323 at 1/200 (2.575 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human white adipose. The section was incubated with ab310323 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. PPAR gamma was immunoprecipitated from 0.35 mg HDLM-2 (human Hodgkin lymphoma) whole cell lysate with ab310323 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab310323 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HDLM-2 (human Hodgkin lymphoma) whole cell lysate Lane 2 : ab310323 IP in HDLM-2 (human Hodgkin lymphoma) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab310323 in HDLM-2 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-PPAR gamma antibody [EPR25862-79] (<a href='/en-us/products/primary-antibodies/ppar-gamma-2-ppar-gamma-antibody-epr25862-79-ab310323'>ab310323</a>) at 1/30 dilution

All lanes:

HDLM-2 (human Hodgkin lymphoma) 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: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Panel A 3T3-L1(mou tissue labeling PPAR gamma with ab310323 at 1/200 (2.575 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on (A) 3T3-L1 cell pellet differentiated for 6 days; no staining on (B) undifferentiated 3T3-L1 cell pellet. The section was incubated with ab310323 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 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-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse brown adipose tissue labelling PPAR gamma with ab310323 at 1/200 (2.575 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse brown adipose. The section was incubated with ab310323 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 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-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse white adipose tissue labelling PPAR gamma with ab310323 at 1/200 (2.575 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse white adipose. The section was incubated with ab310323 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 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-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat white adipose tissue labeling PPAR gamma with ab310323 at 1/200 (2.575 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat white adipose. The section was incubated with ab310323 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 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-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat brown adipose tissue labeling PPAR gamma with ab310323 at 1/200 (2.575 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat brown adipose. The section was incubated with ab310323 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. PPAR gamma was immunoprecipitated from 0.35 mg 3T3-L1 differentiation for 6 days whole cell lysate with ab310323 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab310323 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : 3T3-L1 differentiation for 6 days whole cell lysate Lane 2 : ab310323 IP in 3T3-L1 differentiation for 6 days whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab310323 in 3T3-L1 differentiation for 6 days whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 87 seconds The bands beneath the target band (53kDa) are expected to be degraded target fragments.

All lanes:

Immunoprecipitation - Anti-PPAR gamma antibody [EPR25862-79] (<a href='/en-us/products/primary-antibodies/ppar-gamma-2-ppar-gamma-antibody-epr25862-79-ab310323'>ab310323</a>) at 1/30 dilution

All lanes:

3T3-L1 differentiation for 6 days whole cell lysate at 10 µg

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: 87s

• WB

Supplier Data

Western blot - Anti-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : MOLT-4 (PMID : 7787419). The expression of PPARG is upregulated in response to 3T3-L1 differentiation for 6 days treatment (PMID : 9065481). The bands beneath the target band (53kDa) are expected to be degraded target fragments. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 37 seconds

All lanes:

Western blot - Anti-PPAR gamma antibody [EPR25862-79] (<a href='/en-us/products/primary-antibodies/ppar-gamma-2-ppar-gamma-antibody-epr25862-79-ab310323'>ab310323</a>) at 1/1000 dilution

Lane 1:

HDLM-2 (human Hodgkin lymphoma) whole cell lysate at 50 µg

Lane 2:

Untreated 3T3-L1 (mouse embryonic fibroblast) whole cell lysate at 50 µg

Lane 3:

3T3-L1 differentiation for 6 days whole cell lysate at 50 µg

Lane 4:

MOLT-4 whole cell lysate at 50 µg

Lane 5:

Mouse brown fat tissue lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 53 kDa,58 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-PPAR gamma antibody [EPR25862-79] - BSA and Azide free (AB310324)

This data was developed using ab310323, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody does not cross-react with human PPARD and PPARA. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. Exposure time : 15 seconds

All lanes:

Western blot - Anti-PPAR gamma antibody [EPR25862-79] (<a href='/en-us/products/primary-antibodies/ppar-gamma-2-ppar-gamma-antibody-epr25862-79-ab310323'>ab310323</a>) at 1/1000 dilution

Lane 1:

His-tagged human PPARA recombinant protein at 10 ng

Lane 2:

His-tagged human PPARD recombinant protein at 10 ng

Lane 3:

His-tagged human PPARG recombinant fragment at 10 ng

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: 37 kDa

false

Exposure time: 15s