Anti-MAX antibody [EPR19352] - ChIP Grade

• ChIP-seq

Lab

ChIP-sequencing - Anti-MAX antibody [EPR19352] - ChIP Grade (AB199489)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 8 μg of ab199489 [EPR19352]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. Additional screenshots of mapped reads can be downloaded here.

• IP

Supplier Data

Immunoprecipitation - Anti-MAX antibody [EPR19352] - ChIP Grade (AB199489)

MAX was immunoprecipitated from 0.35 mg of HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate with ab199489 at 1/60 dilution. Western blot was performed from the immunoprecipitate using ab199489 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : 293 whole cell lysate 10μg (Input).

Lane 2 : ab199489 IP in HEK-293 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199489 in HEK-293 whole cell lysate.

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

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-MAX antibody [EPR19352] - ChIP Grade (ab199489)

Predicted band size: 18 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-MAX antibody [EPR19352] - ChIP Grade (AB199489)

MAX was immunoprecipitated from 0.35 mg Mouse brain tissue lysate 10 µg with ab199489 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab199489 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution. Lane 1 : Mouse brain tissue lysate 10 µg

Lane 2 : ab199489 IP in Mouse brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199489 in mouse brain tissue lysate

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

Exposure time : 3 minutes

All lanes:

Immunoprecipitation - Anti-MAX antibody [EPR19352] - ChIP Grade (ab199489)

Predicted band size: 18 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-MAX antibody [EPR19352] - ChIP Grade (AB199489)

MAX was immunoprecipitated from 0.35 mg Rat brain tissue lysate 10 µg with ab199489 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab199489 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution. Lane 1 : Rat brain tissue lysate 10 µg

Lane 2 : ab199489 IP in Rat brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199489 in rat brain tissue lysate

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

Exposure time : 3 minutes

All lanes:

Immunoprecipitation - Anti-MAX antibody [EPR19352] - ChIP Grade (ab199489)

Predicted band size: 18 kDa

false

• WB

Supplier Data

Western blot - Anti-MAX antibody [EPR19352] - ChIP Grade (AB199489)

Blocking/Dilution buffer : 5% NFDM/TBST.

Lanes 1 - 4:

Western blot - Anti-MAX antibody [EPR19352] - ChIP Grade (ab199489) at 1/1000 dilution

Lane 5:

Western blot - Anti-MAX antibody [EPR19352] - ChIP Grade (ab199489) at 1/5000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg

Lane 4:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 10 µg

Lane 5:

Raji (Human Burkitt's lymphoma 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: 18 kDa

Observed band size: 19 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-MAX antibody [EPR19352] - ChIP Grade (AB199489)

Blocking/Dilution buffer : 5% NFDM/TBST.

Recognition of MAX in tissue lysates was very weak and requires optimization.

All lanes:

Western blot - Anti-MAX antibody [EPR19352] - ChIP Grade (ab199489) at 1/1000 dilution

All lanes:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/20000 dilution

Predicted band size: 18 kDa

Observed band size: 19 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-MAX antibody [EPR19352] - ChIP Grade (AB199489)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MAX antibody [EPR19352] - ChIP Grade (ab199489) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) nuclear fraction lysate at 10 µg

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) cytoplasmic fraction 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: 18 kDa

Observed band size: 19 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-MAX antibody [EPR19352] - ChIP Grade (AB199489)

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-MAX antibody [EPR19352] - ChIP Grade (ab199489) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain tissue 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 1/100000 dilution

Predicted band size: 18 kDa

Observed band size: 19 kDa

false