Anti-SAA3 antibody [EPR23102-242] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SAA3 antibody [EPR23102-242] - BSA and Azide free (AB280893)

This data was developed using ab233547, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling SAA3 with ab233547 at 1/50 dilution (9.18 ug/ml), followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SAA3 antibody [EPR23102-242] - BSA and Azide free (AB280893)

This data was developed using ab233547, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling SAA3 with ab233547 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-SAA3 antibody [EPR23102-242] - BSA and Azide free (AB280893)

This data was developed using ab233547, the same antibody clone in a different buffer formulation.

SAA3 was immunoprecipitated from 0.35 mg of Mouse colon tissue lysate with ab233547 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab233547 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse colon tissue lysate 10 ug

Lane 2 : ab233547 IP in Mouse colon tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab233547 in Mouse colon tissue lysate

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

Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-SAA3 antibody [EPR23102-242] (<a href='/en-us/products/primary-antibodies/saa3-antibody-epr23102-242-ab233547'>ab233547</a>)

false

• WB

Lab

Western blot - Anti-SAA3 antibody [EPR23102-242] - BSA and Azide free (AB280893)

This data was developed using ab233547, the same antibody clone in a different buffer formulation.

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

Low expression : kidney and heart (PMID : 29056547)

Exposure time : 92 seconds

All lanes:

Western blot - Anti-SAA3 antibody [EPR23102-242] (<a href='/en-us/products/primary-antibodies/saa3-antibody-epr23102-242-ab233547'>ab233547</a>) at 1/1000 dilution

Lane 1:

Rat colon tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Rat spleen tissue lysate at 20 µg

Lane 4:

Rat kidney tissue lysate at 20 µg

Lane 5:

Rat heart tissue 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 1/100000 dilution

Observed band size: 14 kDa

false

• WB

Lab

Western blot - Anti-SAA3 antibody [EPR23102-242] - BSA and Azide free (AB280893)

This data was developed using ab233547, the same antibody clone in a different buffer formulation.

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

Low expression : kidney and heart (PMID : 29056547)

Exposure time : 92 seconds

All lanes:

Western blot - Anti-SAA3 antibody [EPR23102-242] (<a href='/en-us/products/primary-antibodies/saa3-antibody-epr23102-242-ab233547'>ab233547</a>) at 1/1000 dilution

Lane 1:

Mouse colon tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse heart tissue lysate at 20 µg

Lane 5:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 6:

J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

Lane 7:

NR8383 (rat norvegicus lung macrophage (alveolar)) 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 1/100000 dilution

Observed band size: 14 kDa

false

• WB

Lab

Western blot - Anti-SAA3 antibody [EPR23102-242] - BSA and Azide free (AB280893)

This data was developed using ab233547, the same antibody clone in a different buffer formulation.

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

Exposure time : 26 seconds

All lanes:

Western blot - Anti-SAA3 antibody [EPR23102-242] (<a href='/en-us/products/primary-antibodies/saa3-antibody-epr23102-242-ab233547'>ab233547</a>) at 1/1000 dilution

Lane 1:

His-tagged mouse Saa1 recombinant protein (aa 20-122) at 0.02 µg

Lane 2:

E.coli extracts containing his-tagged mouse Saa2 recombinant protein (aa 20-122) at 20 µL

Lane 3:

His/lif-tagged mouse Saa3 recombinant protein (aa 19-122) at 0.01 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

false