Q1: The ascites-specific adjuvant is effective, but the injection resistance is high and the needle容易脱针. How to deal with this?
Our ascites adjuvant has a different composition from the conventional adjuvants. It is relatively more viscous and can be used with a syringe without a rubber stopper or a release needle. Rubber-free syringe, anti-disengagement syringe
Q2: Does injecting hybridoma cells beyond a few days have any impact?
The instruction manual states that 12 to 18 days after sensitization, hybridoma cells should be injected. Generally, it is fine within 25 days after sensitization. We have done it for up to 30 days, which is still okay. However, 12 to 18 days is the best. If it exceeds 25 days, it is generally not recommended to do it again. You can try your luck and directly inject the cells without adding adjuvants.
Q3: Low or no ascites production
(1) Incorrect sensitization, incorrect use of sensitizers, or a significant time difference between sensitization and hybridoma inoculation, usually 7 to 14 days, depending on the abdominal condition of the mouse. (2) The hybridoma cells or sensitizers were inoculated at incorrect locations, not reaching the abdominal cavity but subcutaneous. (3) The number of hybridoma cells inoculated is too small, too many or the cell condition is poor. Generally, it is advisable not to have around 500,000 cells. (4) It is recommended to use female mice or those that have given birth to prepare ascites.
Q4: Low ascites titer
(1) The hybridoma used to prepare ascites is extremely unstable. It loses its antibody secretion capacity before being injected into mice or within the abdominal cavity. (2) The wrong sensitizer was used when sensitizing the mice; (3) The number of hybridoma cells inoculated is too small, or the cell condition is poor; (4) Due to incorrect injection methods and sites, solid tumors were formed.
Q5: Formation of solid tumors
Two situations may lead to solid tumors: (1) The number of hybridoma cells is too large, or the "toxicity" of hybridoma cells is relatively strong, and the number of hybridomas needs to be appropriately reduced; (2) The hybridoma cells have been contaminated by microorganisms, and the injected hybridoma cells are impure. In this case, the hybridoma cells need to be taken and re-inoculated with 5x antibiotics, and then ascites should be prepared.
Q6: 使用注意事项
腹腔注射点在小鼠左侧或右侧的下腹部，避开肝脏和较低
Q7: 1. The ascites-specific adjuvant is effective, but the injection resistance is high and the needle容易脱针. How to deal with this?
Our ascites adjuvant has a different composition from the conventional adjuvants. It is relatively more viscous and can be used with a syringe without a rubber stopper or a release needle.
Q8: 2. Low or no ascites production
(1) Incorrect sensitization, incorrect use of sensitizers, or a significant time difference between sensitization and hybridoma inoculation, usually 7 to 14 days, depending on the abdominal condition of the mouse;
(2) The hybridoma cells or sensitizers were inoculated at incorrect locations, not reaching the abdominal cavity but subcutaneous.
(3) The number of hybridoma cells inoculated is too small, too many or the cell condition is poor. Generally, it is advisable not to have around 500,000 cells.
(4) It is recommended to use female mice or those that have given birth to prepare ascites.
Q9: 3. Low ascites titer
(1) The hybridoma used to prepare ascites is extremely unstable. It loses its antibody secretion capacity before being injected into mice or once injected into the abdominal cavity.
(2) The wrong sensitizer was used when sensitizing the mice;
(3) The number of hybridoma cells inoculated is too small, or the cell condition is poor;
(4) Due to incorrect injection methods and sites, solid tumors were formed.
Q10: 4. Formation of solid tumors
Two situations may lead to solid tumors:
(1) The number of hybridoma cells is too large, or the hybridoma cells ";" "Toxicity" It is relatively strong and the number of hybridomas needs to be appropriately reduced.
(2) The hybridoma cells have been contaminated by microorganisms, and the injected hybridoma cells are impure. In this case, the hybridoma cells need to be taken and re-inoculated with 5x antibiotics, and then ascites should be prepared.
Q11: 5. Usage precautions
The intraperitoneal injection site was located in the lower abdomen on the left or right side of the mouse, avoiding the liver and the lower and middle abdominal bladder, and penetrated the peritoneum along the Angle of the leg. When the 1ml syringe needle enters about one-third, there is a feeling of being pierced through.
Q12: 腹水专用佐剂效果虽好，但是注射阻力大易脱针，怎么处理
我们的腹水专用佐剂和常规的佐剂成分不同，会相对粘稠一些，可以使用无橡胶塞或者放脱针的注射器。
Q13: 注射杂交瘤细胞，超过几天有没有影响
说明书上说致敏后12-18天，注射杂交瘤细胞。一般致敏后25内都是可以的，最多我们做过30天的，也还可以，但是12-18天是最好的，超过25天的话一般不建议再打，可以试试运气，直接打细胞，不需要加免佐剂。
Q14: 腹水少或无腹水
（1）致敏不正确，不正确的使用致敏剂，或者致敏时间与接种杂交瘤的时间相差太久，一般是7-14天，具体看小鼠的腹部情况；
（2）杂交瘤细胞或者致敏剂的接种位置不正确，没有打到腹腔，而是打到了皮下；
（3）接种的杂交瘤细胞数量太少，太多或者细胞状态不好。一般不50万个细胞左右为宜；
（4）建议使用母鼠或者生育过的母鼠制备腹水。
Q15: 腹水效价低
（1）制备腹水的杂交瘤极不稳定，打到小鼠身上之前就失去抗体分泌能力或者打到腹腔内就失去了分泌能力；
（2）致敏小鼠时采用了错误的致敏剂
（3）接种的杂交瘤细胞数量太少，或者细胞状态不好
（4）由于注射方式和部位错误，形成了实体瘤。
Q16: 形成实体瘤
两种情况可能出现实体瘤：（1）杂交瘤细胞量太大，或者杂交瘤细胞“毒性”比较强，需要适当减少杂交瘤数量；
（2）杂交瘤细胞有被微生物污染，注射的杂交瘤细胞不纯，这个情况需要取杂交瘤细胞加5x抗生素重新进行亚克隆化，然后在进行腹水制备。
Q17: Usage precautions.
The intraperitoneal injection site was located in the lower abdomen on the left or right side of the mouse, avoiding the liver and the lower and middle abdominal bladder, and penetrated the peritoneum along the Angle of the leg. When the 1ml syringe needle enters about one-third, there is a feeling of being pierced through.
Q18: What is the material of the needle in the adjuvant emulsifier? Worried about residues in the reagent from the vibrating needle.
No residues have been detected in stainless steel and copper for the time being, nor have any feedback been received regarding any residues
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