Anti-CD31 antibody [EPR17260-254] - BSA and Azide free

• IHC-tissue clearing

Supplier Data

IHC-tissue clearing - Anti-CD31 antibody [EPR17260-254] - BSA and Azide free (AB271994)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213175).

Anti-CD31 ab213175 was used with Tissue clearing kit – CUBIC (ab316246) and 3D Tissue Staining Kit – CUBIC (ab316248) to penetrate, stain and clear a whole mouse brain.

Learn more about tissue clearing kits, reagents, and protocols designed to make it easier to stain whole brains and get more data from each valuable tissue sample.

For a whole mouse brain, we recommend starting with 10 ug of ab213175 and using a Fab fragment secondary antibody with 6.67 µg  to create an antibody complex before 3D staining (see protocol for details). No additive was used during the staining process.

The sample was imaged using a light-sheet microscopy.

• IP

Supplier Data

Immunoprecipitation - Anti-CD31 antibody [EPR17260-254] - BSA and Azide free (AB271994)

CD31 was immunoprecipitated from 1 mg of bEnd.3 (Mouse brain endothelioma cell line) whole cell lysate with ab213175 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab213175 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1 : bEnd.3 whole cell lysate 10μg (Input). Lane 2 : ab213175 IP in bEnd.3 whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab213175 in bEnd.3 whole cell lysate.

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

Exposure time : 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213175).

All lanes:

Immunoprecipitation - Anti-CD31 antibody [EPR17260-254] (<a href='/en-us/products/primary-antibodies/cd31-antibody-epr17260-254-ab213175'>ab213175</a>)

Predicted band size: 82 kDa

false

• ELISA

Supplier Data

ELISA - Anti-CD31 antibody [EPR17260-254] - BSA and Azide free (AB271994)

Example of mouse CD31 standard curve in 1X Cell Extraction Buffer PTR using recombinant CD31 antibody (ab213175) and a recombinant monoclonal antibody. Background-subtracted data values (mean +/- SD) are graphed.

For a matched antibody pair set, see Mouse CD31 Matched Antibody Pair Kit (ab212065).

For a complete ELISA kit, see Mouse CD31 ELISA kit (ab204527).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213175).

• WB

Supplier Data

Western blot - Anti-CD31 antibody [EPR17260-254] - BSA and Azide free (AB271994)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213175).

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1-3 : 10 seconds; Lane 4/5 : 1 second.

The expression MW observed is consistent with what has been described in the literature (PMID : 18388311; 12433657).

Lanes 1 - 4:

Western blot - Anti-CD31 antibody [EPR17260-254] (<a href='/en-us/products/primary-antibodies/cd31-antibody-epr17260-254-ab213175'>ab213175</a>) at 1/1000 dilution

Lane 5:

Western blot - Anti-CD31 antibody [EPR17260-254] (<a href='/en-us/products/primary-antibodies/cd31-antibody-epr17260-254-ab213175'>ab213175</a>) at 1/2000 dilution

Lane 1:

Mouse platelet lysate at 10 µg

Lane 2:

Rat platelet lysate at 10 µg

Lane 3:

Rat lung lysate at 10 µg

Lane 4:

Mouse lung lysate at 10 µg

Lane 5:

bEnd.3 (Mouse brain endothelioma cell line) whole cell lysate at 10 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

Predicted band size: 82 kDa

Observed band size: 110-130 kDa

false

• WB

Supplier Data

Western blot - Anti-CD31 antibody [EPR17260-254] - BSA and Azide free (AB271994)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213175).

Blocking/Dilution buffer : 5% NFDM/TBST.

Lane 1:

Western blot - Anti-CD31 antibody [EPR17260-254] (<a href='/en-us/products/primary-antibodies/cd31-antibody-epr17260-254-ab213175'>ab213175</a>) at 1/20000 dilution

Lane 2:

Western blot - Anti-CD31 antibody [EPR17260-254] (<a href='/en-us/products/primary-antibodies/cd31-antibody-epr17260-254-ab213175'>ab213175</a>) at 1/100000 dilution

All lanes:

Mouse CD31 active protein 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 1/20000 dilution

Predicted band size: 82 kDa

Observed band size: 100 kDa

false

Exposure time: 3min