Anti-BNIP3 antibody [EPR4034] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] - BSA and Azide free (AB219609)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human renal adenocarcinoma tissue labelling BNIP3 with purified ab109362 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109362).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-BNIP3 antibody [EPR4034] - BSA and Azide free (AB219609)

Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y cells labelling BNIP3 with unpurified ab109362 at a 1/100 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109362).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] - BSA and Azide free (AB219609)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling BNIP3 with unpurified ab109362 at a 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109362).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-BNIP3 antibody [EPR4034] - BSA and Azide free (AB219609)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109362).

Lane 1 : Wild-type HAP1 whole cell lysate (40 μg)
Lane 2 : BNIP3 knockout HAP1 whole cell lysate (40 μg)
Lane 3 : SHSY5Y whole cell lysate (40 μg)
Lane 4 : A431 whole cell lysate (40 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109362 observed at 25 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab109362 was shown to recognize BNIP3 in wild-type HAP1 cells as signal was lost at the expected MW in BNIP3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and BNIP3 knockout samples were subjected to SDS-PAGE. ab109362 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-BNIP3 antibody [EPR4034] (<a href='/en-us/products/primary-antibodies/bnip3-antibody-epr4034-ab109362'>ab109362</a>)

Predicted band size: 21 kDa

false

• WB

Supplier Data

Western blot - Anti-BNIP3 antibody [EPR4034] - BSA and Azide free (AB219609)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109362). The observed band sizes are consistent with publications. Bands around 30 and 60kDa should be monomer and dimer, respectively. Other bands should be different migrating forms (PMID : 15375294, PMID : 36396625, PMID : 9396766, PMID : 36396625, PMID : 33459136 etc.). 1% SDS hot lysis method could help to get stronger monomer forms. The protocol used for this test is available under the Support & downloads section

All lanes:

Western blot - Anti-BNIP3 antibody [EPR4034] (<a href='/en-us/products/primary-antibodies/bnip3-antibody-epr4034-ab109362'>ab109362</a>) at 1/1000 dilution

Lane 1:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate 20ug, prepared by RIPA method at 20 µg

Lane 2:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate 20ug, prepared by 1% SDS hot method 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

false

Exposure time: 156s