Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸

Order now and get it on Wednesday March 10, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y244] to CrkL
Suitable for: WB, IHC-P, ICC, IP
Knockout validated
Reacts with: Human

Product name

Anti-CrkL antibody [Y244]
See all CrkL primary antibodies
Description

Rabbit monoclonal [Y244] to CrkL
Host species

Rabbit
Specificity

ab32018 recognises CrkL (Crk-like)protein.

Tested Applications & Species

Application	Species
ICC	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human CrkL aa 250-350 (C terminal). The exact sequence is proprietary.
Positive control
- HeLa and K562 cell lysates, human metastatic lung carcinoma
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

IgG fraction
Clonality

Monoclonal
Clone number

Y244
Isotype

IgG
Research areas

Western blot - Anti-CrkL antibody [Y244] (ab32018)

All lanes : Anti-CrkL antibody [Y244] (ab32018) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CRKL knockout HeLa cell lysate
Lane 3 : K-562 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 33 kDa
Observed band size: 37 kDa
why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab32018 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.

ab32018 Anti-CrkL antibody [Y244] was shown to specifically react with CrkL in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265993 (knockout cell lysate ab257397) was used. Wild-type and CrkL knockout samples were subjected to SDS-PAGE. ab32018 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry - Anti-CrkL antibody [Y244] (ab32018)

Immunocytochemistry analysis of K-562 (human chronic myelogenous leukemia lymphoblast) labeling CrkL with purified ab32018 at 1/100 dilution (10 µg/mL). Cells were fixed with 100% methanol. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/mL) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control: PBS instead of the primary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CrkL antibody [Y244] (ab32018)

ab32018, at a 1/50 dilution, staining human Crk in metastatic lung carcinoma by Immunohistochemistry, in paraffin embedded tissue

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-CrkL antibody [Y244] (ab32018)

Purified ab32018 at 1/50 dilution (2µg) immunoprecipitating CrkL in K-562 whole cell lysate.
Lane 1 (input): K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg
Lane 2 (+): ab32018 + K-562 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32018 in K-562 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 37 kDa
Western blot - Anti-CrkL antibody [Y244] (ab32018)

Anti-CrkL antibody [Y244] (ab32018) at 1/500 dilution + K562 cell lysate

Predicted band size: 33 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Anti-CrkL antibody [Y244] (ab32018)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com