Anti-Complement factor 8 beta/C8B antibody [EPR23764-1]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (AB278045)

Immunohistochemical analysis of paraffin-embedded human thyroid tissue labeling Complement factor 8 beta/C8B with ab278045 at 1/1000 (0.533 μg/ml) dilution followed by a ready to use secondary antibody. Positive staining on the blood plasma in human thyroid (PMID : 9476128). The section was incubated with ab278045 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 secondary antibody.

Heat mediated antigen retrieval 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-Complement factor 8 beta/C8B antibody [EPR23764-1] (AB278045)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Complement factor 8 beta/C8B with ab278045 at 1/1000 (0.533 μg/ml) dilution followed by a ready to use secondary antibody. The section was incubated with ab278045 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Negative control : no staining on human cerebrum.

Secondary antibody only control : Secondary antibody is a ready to use secondary antibody.

Heat mediated antigen retrieval 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-Complement factor 8 beta/C8B antibody [EPR23764-1] (AB278045)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Complement factor 8 beta/C8B with ab278045 at 1/1000 (0.533 μg/ml) dilution followed by a ready to use secondary antibody. Cytoplasmic staining on human liver. The section was incubated with ab278045 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 secondary antibody).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (AB278045)

Complement factor 8 beta/C8B was immunoprecipitated from 0.35 mg human kidney tissue lysate with ab278045 at 1/30 dilution (2μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab278045 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human kidney tissue lysate 10 μg

Lane 2 : ab278045 IP in human kidney tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab278045 in human kidney tissue lysate

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

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (ab278045)

Predicted band size: 67 kDa

Observed band size: 67 kDa

false

• WB

Lab

Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (AB278045)

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

The observed MW is consistent with what has been described in the literature (PMID : 21750743).

Negative : human brain (PMID : 16483657, PMID : 27005612).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (ab278045) at 1/1000 dilution

Lane 1:

Human kidney tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human large intestine tissue lysate at 20 µg

Lane 4:

Human brain tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 67 kDa

Observed band size: 67 kDa

false

• WB

Lab

Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (AB278045)

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

The observed MW is consistent with what has been described in the literature (PMID : 21750743).

Exposure time : 15 seconds.

All lanes:

Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (ab278045) at 1/1000 dilution

All lanes:

Human serum at 20 µL

Secondary

All lanes:

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

Predicted band size: 67 kDa

Observed band size: 67 kDa

false

• WB

Lab

Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (AB278045)

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

This antibody has no cross reaction with human C8A and C9.

Sample loaded onto lane 1 was purified protein extracted from HEK293 expressing C8B, which contains a LIF-tag (40 kDa).

C8A and C9 recombinant proteins were expressed in E.coil expression systems.

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (ab278045) at 1/1000 dilution

Lane 1:

LIF-tagged human C8B recombinant protein, 30 ng

Lane 2:

E.coli extracts containing his-tagged human C8A recombinant protein, 30 ng

Lane 3:

E.coli extracts containing his-tagged human C9 recombinant protein, 30 ng

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 67 kDa

false