Anti-alpha 2 Macroglobulin antibody [EPR25958-49] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 2 Macroglobulin antibody [EPR25958-49] - BSA and Azide free (AB316102)

This data was developed using ab316101, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) Non-perfused fixed rat kidney (B) Perfused fixed rat kidney tissue labeling alpha 2 Macroglobulin with ab316101 at 1/500 (1.032 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on plasma of (A) Non-perfused fixed rat kidney, negative staining on (B) Perfused fixed rat kidney. The section was incubated with ab316101 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 2 Macroglobulin antibody [EPR25958-49] - BSA and Azide free (AB316102)

This data was developed using ab316101, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) Non-perfused fixed rat liver (B) Perfused fixed rat liver tissue labeling alpha 2 Macroglobulin with ab316101 at 1/500 (1.032 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on plasma of (A) Non-perfused fixed rat liver, negative staining on (B) Perfused fixed rat liver. The section was incubated with ab316101 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 2 Macroglobulin antibody [EPR25958-49] - BSA and Azide free (AB316102)

This data was developed using ab316101, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Non-perfused fixed rat E20 liver tissue labeling alpha 2 Macroglobulin with ab316101 at 1/500 (1.032 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on non-perfused fixed rat E20 liver. The section was incubated with ab316101 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.

• IP

Supplier Data

Immunoprecipitation - Anti-alpha 2 Macroglobulin antibody [EPR25958-49] - BSA and Azide free (AB316102)

This data was developed using ab316101, the same antibody clone in a different buffer formulation.

alpha 2 Macroglobulin was immunoprecipitated from 0.35 mg Rat E20 liver tissue lysate with ab316101 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316101 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-alpha 2 Macroglobulin antibody [EPR25958-49] (<a href='/ko/products/primary-antibodies/alpha-2-macroglobulin-antibody-epr25958-49-ab316101'>ab316101</a>) at 1/1000 dilution

Lane 1:

Rat E20 liver tissue lysate

Lane 2:

<a href='/ko/products/primary-antibodies/alpha-2-macroglobulin-antibody-epr25958-49-ab316101'>ab316101</a> at 1/30 IP in Rat E20 liver tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/ko/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/ko/products/primary-antibodies/alpha-2-macroglobulin-antibody-epr25958-49-ab316101'>ab316101</a> in rat E20 liver tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-alpha 2 Macroglobulin antibody [EPR25958-49] - BSA and Azide free (AB316102)

This data was developed using ab316101, the same antibody clone in a different buffer formulation.

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

Low expression : liver, heart (PMID : 2450021).

The molecular weight observed is consistent with what has been described in the literature (PMID : 31642940).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-alpha 2 Macroglobulin antibody [EPR25958-49] (<a href='/ko/products/primary-antibodies/alpha-2-macroglobulin-antibody-epr25958-49-ab316101'>ab316101</a>) at 1/1000 dilution

Lane 1:

Rat plasma at 20 µg

Lane 2:

Rat serum at 20 µg

Lane 3:

Rat eyeball tissue lysate at 20 µg

Lane 4:

Rat E20 liver tissue lysate at 20 µg

Lane 5:

Rat E17 liver tissue lysate at 20 µg

Lane 6:

Rat liver tissue lysate at 20 µg

Lane 7:

Rat heart 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: 180 kDa,125 kDa,90 kDa,124 kDa

false

Exposure time: 15s