Human HSPB8 (Hsp22) knockout HeLa cell line
- Advanced Validation
- What is this?
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HSPB8 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 2 bp deletion in exon 1.
View Alternative Names
Alpha-crystallin C chain, CMT2L, CRYAC, Charcot Marie Tooth disease axonal type 2L, Charcot Marie Tooth disease spinal, DHMN 2, E2-induced gene 1 protein, E2IG1, HMN 2, HMN2A, HSB8, HSPB8_HUMAN, Heat shock 22kDa protein 8, Heat shock 27kDa protein 8, Heat shock protein 22, Heat shock protein beta-8, Hereditary motor neuropathy distal, OTTHUMP00000239768, Protein kinase H11, Small stress protein-like protein HSP22, Spinal muscular atrophy distal adult autosomal dominant
- WB
Lab
Western blot - Human HSPB8 (Hsp22) knockout HeLa cell line (AB265112)
Lanes 1 - 2 : Merged signal (red and green). Green - ab151552 observed at 212 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab151552 was shown to react with Hsp22/HSPB8 in wild-type HeLa cells in Western blot with loss of signal observed in HSPB8 knockout cell line ab265112 (HSPB8 knockout cell lysate ab257468). Wild-type HeLa and HSPB8 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab151552 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-Hsp22/HSPB8 antibody [EPR9714] (<a href='/en-us/products/primary-antibodies/hsp22-hspb8-antibody-epr9714-ab151552'>ab151552</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
HSPB8 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
Western blot - Human HSPB8 (Hsp22) knockout HeLa cell line (ab265112)
Predicted band size: 21 kDa
Observed band size: 21 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human HSPB8 (Hsp22) knockout HeLa cell line (AB265112)
Allele-1 : 2 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human HSPB8 (Hsp22) knockout HeLa cell line (AB265112)
Allele-2 : 1 bp deletion in exon 1.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Hsp22 plays a critical role in maintaining protein homeostasis as part of a larger chaperone complex. It forms hetero-oligomers with other small heat shock proteins like HSPB1 and HSPB6 enhancing its protein-protective functions. These interactions allow Hsp22 to stabilize cytoskeletal elements and partake in autophagic pathways highlighting its importance during cellular stress conditions.
Pathways
Hsp22 integrates into cellular processes of autophagy and proteostasis. It participates in the chaperone-assisted selective autophagy (CASA) pathway. Here it associates with the cochaperone BAG3 facilitating the degradation of damaged proteins. Hsp22 also plays a role in the heat shock response pathway cooperating with other heat shock proteins like HSP70 to protect cells from damage induced by increased temperatures.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB151552
Anti-Hsp22/HSPB8 antibody [EPR9714]
RabMAb|Recombinant|KO Validated|Trial Size
![Western blot - Anti-Hsp22/HSPB8 antibody [EPR9714] (AB151552)](https://content.abcam.com/products/images/hsp22-hspb8-antibody-epr9714-ab151552--western-blot-img150544.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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