Anti-BAF170 antibody [EPR23066-21] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-BAF170 antibody [EPR23066-21] - BSA and Azide free (AB263871)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized A549 (human lung carcinoma epithelial cell) cells labelling BAF170 with ab243634 at 1/500 (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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243634).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BAF170 antibody [EPR23066-21] - BSA and Azide free (AB263871)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma epithelial cell) cells labeling BAF170 with ab243634 at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in A549 cell line. The nuclear counter stain is DAPI (Blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red).

PBS only control : Used PBS instead of primary antibody, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243634).

• IP

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Immunoprecipitation - Anti-BAF170 antibody [EPR23066-21] - BSA and Azide free (AB263871)

BAF170 was immunoprecipitated from 0.35 mg K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab243634 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab243634 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10ug

Lane 2 : ab243634 IP in K-562 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab243634 in K-562 whole cell lysate

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

Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243634).

All lanes:

Immunoprecipitation - Anti-BAF170 antibody [EPR23066-21] (<a href='/en-us/products/primary-antibodies/baf170-antibody-epr23066-21-ab243634'>ab243634</a>)

Predicted band size: 133 kDa

Observed band size: 170 kDa

false

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-BAF170 antibody [EPR23066-21] - BSA and Azide free (AB263871)

Intracellular flow cytometric analysis of Neuro-2a (mouse neuroblastoma neuroblast) cells labelling BAF170 with ab243649 at 1/500 (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). Goat anti rabbit IgG (Alexa Fluor^®647, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243634).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BAF170 antibody [EPR23066-21] - BSA and Azide free (AB263871)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labeling BAF170 with ab243634 at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in Neuro-2a cell line. The nuclear counter stain is DAPI (Blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red).

PBS only control : Used PBS instead of primary antibody, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243634).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BAF170 antibody [EPR23066-21] - BSA and Azide free (AB263871)

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling BAF170 with ab243634 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse hippocampus (PMID : 27392482) The section was incubated with ab243634 for 15 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243634).

• WB

Lab

Western blot - Anti-BAF170 antibody [EPR23066-21] - BSA and Azide free (AB263871)

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

This antibody also recognizes weak unidentifiable bands below 100kDa.

Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243634).

All lanes:

Western blot - Anti-BAF170 antibody [EPR23066-21] (<a href='/en-us/products/primary-antibodies/baf170-antibody-epr23066-21-ab243634'>ab243634</a>) at 1/1000 dilution

Lane 1:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Lane 2:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 5:

C2C12 (mouse myoblasts myoblast), whole cell lysate at 20 µg

Lane 6:

Mouse brain tissue lysate at 10 µg

Lane 7:

Mouse lung tissue lysate at 10 µg

Lane 8:

Rat brain tissue lysate at 10 µg

Lane 9:

Rat liver tissue 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 1/50000 dilution

Predicted band size: 133 kDa

Observed band size: 170 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-BAF170 antibody [EPR23066-21] - BSA and Azide free (AB263871)

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

This antibody also recognizes weak unidentifiable bands below 100kDa.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243634).

All lanes:

Western blot - Anti-BAF170 antibody [EPR23066-21] (<a href='/en-us/products/primary-antibodies/baf170-antibody-epr23066-21-ab243634'>ab243634</a>) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

Lane 2:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell at 10 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast), 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 1/50000 dilution

Predicted band size: 133 kDa

Observed band size: 170 kDa

false

Exposure time: 3s