Anti-CISH/CIS antibody [EPR25357-36] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-CISH/CIS antibody [EPR25357-36] - BSA and Azide free (AB308501)

This data was developed using ab308500, the same antibody clone in a different buffer formulation.

CISH/CIS was immunoprecipitated from 0.35 mg No-GFP-CD16.NK-92 (human malignant non-Hodgkin's lymphoma natural killer cell) whole cell lysate with ab308500 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308500 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : No-GFP-CD16.NK-92 (human malignant non-Hodgkin's lymphoma natural killer cell) whole cell lysate
Lane 2 : ab308500 IP in No-GFP-CD16.NK-92 (human malignant non-Hodgkin's lymphoma natural killer cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308500 in No-GFP-CD16.NK-92 whole cell lysate

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

Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-CISH/CIS antibody [EPR25357-36] (<a href='/en-us/products/primary-antibodies/cish-cis-antibody-epr25357-36-ab308500'>ab308500</a>) at 1/1000 dilution

All lanes:

No-GFP-CD16.NK-92 (human malignant non-Hodgkin's lymphoma natural killer cell) whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 32 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CISH/CIS antibody [EPR25357-36] - BSA and Azide free (AB308501)

This data was developed using ab308500, the same antibody clone in a different buffer formulation.  Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : Raji (PMID : 11032736), HL-60 (PMID : 11032736). The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 9774439). The expression of CISH/CIS is upregulated in response to IL-2 treatment (PMID : 7796808). This blot was developed using a higher sensitivity ECL substrate. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-CISH/CIS antibody [EPR25357-36] (<a href='/en-us/products/primary-antibodies/cish-cis-antibody-epr25357-36-ab308500'>ab308500</a>) at 1/1000 dilution

Lane 1:

Untreated No-GFP-CD16.NK-92 (human malignant non-Hodgkins lymphoma natural killer cell) whole cell lysate at 20 µg

Lane 2:

No-GFP-CD16.NK-92 treated with /mL IL-2 for 6 hours whole cell lysate at 20 µg

Lane 3:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 4:

HDLM-2 (human Hodgkin lymphoma) whole cell lysate at 20 µg

Lane 5:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 6:

HL-60 (human Acute Promyelocytic Leukemia promyeloblast) 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 1/100000 dilution

Predicted band size: 28 kDa

Observed band size: 32 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CISH/CIS antibody [EPR25357-36] - BSA and Azide free (AB308501)

This data was developed using 308500, the same antibody clone in a different buffer formulation.  Blocking and diluting buffer and concentration : 5% NFDM/TBST This blot was developed using a higher sensitivity ECL substrate. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 9774439). Exposure time : 180 seconds

All lanes:

Western blot - Anti-CISH/CIS antibody [EPR25357-36] (<a href='/en-us/products/primary-antibodies/cish-cis-antibody-epr25357-36-ab308500'>ab308500</a>) at 1/1000 dilution

All lanes:

WEHI-3 (Mouse leukemia lymphoblast) 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 1/100000 dilution

Predicted band size: 28 kDa

Observed band size: 32 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CISH/CIS antibody [EPR25357-36] - BSA and Azide free (AB308501)

This data was developed using 308500, the same antibody clone in a different buffer formulation.  Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : brain (PMID : 7796808). This blot was developed using a higher sensitivity ECL substrate. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 9774439). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-CISH/CIS antibody [EPR25357-36] (<a href='/en-us/products/primary-antibodies/cish-cis-antibody-epr25357-36-ab308500'>ab308500</a>) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse 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: 28 kDa

Observed band size: 32 kDa

false

Exposure time: 180s