Anti-C4B-d / C4B-b antibody [EPR11203-27]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-C4B-d / C4B-b antibody [EPR11203-27] (AB181241)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling C4b with purified ab181241 at 1/100 dilution (10 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C4B-d / C4B-b antibody [EPR11203-27] (AB181241)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling C4b with purified ab181241 at 1/75 dilution (9.53 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Unknown

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] (AB181241)

The molecular weights observed are consistent with what have been described in the literature (PMID : 15632414, 31178241 and 30210498).

All lanes:

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] (ab181241) at 1/10000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

Human plasma lysate at 15 µg

Lane 3:

Human serum lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 193 kDa

Observed band size: 200 kDa,70 kDa,93 kDa

false

• WB

Lab

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] (AB181241)

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

Lanes 1 - 5:

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] (ab181241) at 1/1000 dilution

Lanes 6 - 10:

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (<a href='/en-us/products/primary-antibodies/6x-his-tag-antibody-epr20547-chip-grade-ab213204'>ab213204</a>) at 1/5000 dilution

Lanes 1 and 6:

293T cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

Lanes 2 and 7:

293T cells transfected with a human C4d-A (Uniprot: P0C0L4 aa957-1336) expression vector containing a His-tag whole cell lysate at 20 µg

Lanes 3 and 8:

293T cells transfected with a human C4d-B (Uniprot: P0C0L5 aa957-1336) expression vector containing a His-tag whole cell lysate at 20 µg

Lanes 4 and 9:

293T cells transfected with a human C4-A alpha chain (Uniprot: P0C0L4 aa680- 1446) expression vector containing a His-tag whole cell lysate at 20 µg

Lanes 5 and 10:

293T cells transfected with a human C4b-A (Uniprot: P0C0L4 aa757-1446) expression vector containing a His-tag 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 1/20000 dilution

Observed band size: 45-100 kDa

false

Exposure time: 1s

• WB

Unknown

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] (AB181241)

The molecular weights observed are consistent with what have been described in the literature (PMID : 15632414, 31178241 and 30210498).

All lanes:

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] (ab181241) at 1/10000 dilution

Lane 1:

Human liver lysate at 20 µg

Lane 2:

Human heart lysate at 20 µg

Lane 3:

Human testis lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 193 kDa

Observed band size: 200 kDa,70 kDa,93 kDa

false